biohimija cheloveka t.1
DESCRIPTION
Р.Марри Д.Греннер П.МеЙес В.Родуэпп БИОХИМИЯ ЧЕЛОВЕКА в 2-х томах Том 1TRANSCRIPT
HARPER'S BIOCHEMISTRY Twenty-first Edition
Robert . Murray, D, PhD Professor of Biochemistry University of Toronto
Daryl . Granner, MD Professor and Chairman Department of Molecular .Physiology and Biophysics Professor of Medicine Vanderbilt University Nashville, Tennessee
Peter A.Mayes, PhD, DSc Reader in Biochemistry Royal Veterinary College University of London
Vlctor W. Rodwell, PhD Professor of Biochemistry Purdue University West Lafayette, Indiana
APPLETON & LANGE Norwalk, Connecticut/San Mateo, California
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«» 1993
577.2 20.070
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63 : 2- . . 1. . .: - .: , 1993. - 384 ., .
ISBN 5-03-001774-7
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Freifelder D. Physical Biochemistry: Applications to Biochemi stry and Molecular Biology, Freeman, 1982.
Fruton J. S. Molecules and Life: Historical Essays the Inter play of Chemistry and Biology, Wiley-Interscience, 1972.
Weinberg R. . molecu1es oflife, Sci. m. (Oct.), 1985,253, 48. ( : .)
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versity.)
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Barrett G. . Chcrnistry and Biochernistry of the Arnino Acids, Chaprnan and , 1985.
. G. Tools· of Biochernistry, Wiley, 1977. Friedman . Chernistry and Biochernisrty of the Sulfhydryl
Group in Arnino Acids. Peptides, and Proteins, Pergamon Press, 1973.
Greenstein J. ., Winitz . Chernictry of the Arnino Acids, 3 vols, Wiley, 1961.
Heftman . Chrornatography: Laboratory Handbook of Chrornatographic and Electrophoretic Methods, 3rd ed., Van Nostrand, 1975. .
Touchstone J. . Practice of Thin Layer Chrornatography, Wi ley-Interscience, 1978.
Zweig G., Sherma J. Handbook of Chromatography, 2 vols, CRC Press, 1972.
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stry, Purdue University.)
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. Staphylococcus aureus V8 Glu-, , - . Glu Lys . CNBr-.
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.
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r
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nmu 41
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, , . L- , ;
. ,
() (. 45).
Cantor R., Schimmel . R. Biophysical Chemistry, Part 1: The Conformation of Macromolecules, Freeman, 1980. [ : ., ' . .-: . 1984.]
Chin . ., Wold F. Separation of peptides phosph6cellu lose and other ceIlulose ion exchangers. Methods Enzymol, 1977, 47, 204.
, . G. The Tools of Biochemistry, Wiley, 1977. Craig L. , Cowburn D., Bleich . Methods of the study of
small polypeptide hormones and antibiotics in solution, . Rev. Biochem., 1975. 44, 509.
Dayhojj . (ed.) Atlas ofProtein Sequence and Structure, Vol. 5, National Biomedical Research Foundation, Washing ton, , 1972, Suppl. 1. 1973; Suppl. 2, 1976; Suppl. 3, 1979.
Hash J. . (ed.) Antibiotics. I: Methods in Enzymology, Vol. 43, Academic Press. 1975.
Heftman . Chromatography: Laboratory Handbook of
Chromatographic and Electrophoretic Methods, 3rd ed., Nostrand, 1975.
Jauregu;-Ade/l J., Mart; J. Acidic cleavage of the aspartyl prollne bond and the limitations of the reaction, Anal. Biochem., 1975,69,468.
Mahoney W. , Hermodson . . High-yield cleavage of tryp tophanyl peptide bonds o-iodosobenzoic acid, Bioche mistry, 1979, 18, 3810.
Mahoney W. , Smith . ; Hermodson . . Fragmentation of proteins with o-iodosobenzoic acid: Chemical mecha nism and identification of o-iodosobenzoic acid as reacti ve contaminant that modifies tyrosyl residues. Biochemi stry, 1981, 20. 443.
Marglin ., Merrifield R. . Chemical synthesis ofpeptides and proteins, . Rev. Biochem .. 1970, 39, 841.
Needelman S. . (ed.) Protein Sequence Determination, Sprin ger-Verlag, 1970.
Patthy L., Smith . L. Reversible modification of arginine res idues: Application to sequence studies restriction of tryptic hydrolysis to Iysine residues, J. Biol. Chem., 1975, 250,557.
Pearson J. D. et al. Reversed-phase supports for the resolution of large denatured protein fragments. J. Chromatogr., 1981, 207, 325.
Regnier . F., Gooding . High performance liquid chroma tography of proteins, Anal. Biochem .• 1980, 103. 1.
Snyder S. ., Innes R. . Peptide neurotransmitters. . Rev. Biochem., 1979. 48, 755.
Stewart J. ., Young J. D. Solid Phase Peptide Synthcsis. Free . 1969.
Storm D. R., Rosenthal S., Swanson . . Polymyxin antibio tics, . Rev. Biochem., 1977, 46, 723.
Zweig G., Sherma J. Handbook of Chromatography, 2 vols, CRC Press. 1972.
5
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, , , , .
, , 1907-1908 . . (. 5.1).
.l 5.1. ildl1 6C.1KOB. t'<l
:\10'-=
.
.
70-80%- ,
.
J.I
.
Gly. Ala. Pro
. , ,
. ,
, , ,
.
( ), . 1 3- 4. . ,
, . , 1 , ,·
. . ,
.
, (. 5.2). (),
,
(. 7).
, -. . l 5.2. tl
. .),
(), tl ( l), ()
. , .
; , 8,6 « » ,-, 2-, 13- -. ;
, , ,
(. 26). , .
,
, , , , F . , .
t , (.
). ,
,
. , , ,
«
» , ,
.
L-- . ,
, :
.
) ( , , . . ).
- ,
}'
(. 5.1). , .
44 5
/ .......... / ......... /'N/ N 1
~
. 5.1. ;
C-N.
5.1 ().
, , . . 5.2, , ,
,
.
, .
«»
( ),
. 5.2. . , , . -, -R- . , - - , ( ). , - , · ( , ) . - 0,36 . " ( ). ( , ulig L., Corey R. , Branson . R.: The structure of proteins. Two hydrogen-bonded helical figur tions of the polypeptide chain. Proc. Natl. Acad. Sc;. USA
.1951:37:205.)
. , . ( S-S) J3- ( S-S ) , ;
(. . 4.10).
tl
1) , ; 2) ,
; 3) ,
, .
, . .
(. . «u-) «
J3-»).
.
, . · .
,
. , + 1, - 1. , , .
(, + -) , ,
.
"
,
, .
.
, 1930- ., - 0,5--
0--
. 5.3. " -.
O,5r
. 5.4. - . R - , . ", , " . ( Stryer L. Biochemistry, 2nd ed., Freemal1, 1981. : . I:). 3- .- : . 1985.)
0,55 . , .
(. 5.2). m , , -
- (. 5.3). R- -
(. 5.4). 3,6 , 0,54 , 0,5-{),55 , . , , 0,15 , . '(f (. 5.5).
1. - ,
, ,
.
2. . -.
3. ,
" ( ).
4. - ,
.
5. L- , , - .
46 5
R
. 5.5. -. - ~ R~ ~ ~ ( ). ( Haggis . . et l., Introduetion to
Molccul~r Biology. Wilcy. 1964.)
-,
- . (
,
N - ), R-,
- (. 5.3).
-
- - ( , -).
-
, - (. 5.6). ,
5.3. - -
Cys l
Val
(
N- ), LI
- ( . 5.6). ,
- ( ).
---
-
--- -
. 5.6. .l -. J'. ,10 NH- . (R) . clTOMbI , - , -
. ( Stryer L. Biochemistry. 2nd cd .• Free- , 1981. .)
: 47
. S.7. .n - /l. ~ Jl 124 . . -u- , . . (. 8) POJ-4• ( Kartha G., J., Harker .: Tertiary structure ofribo nuclcase. Nature 1967:213:862.) -
.
. . 5.7 ,
~-CTpYKTypa . . -, - (. 5.7).
-
, ' ,
~-CTpYKTypa ,
. . 5.7.
()
, ,
. . 5.7, . « » :
,
. , - ,
- -.
, (. 4). (
) , .
.
(-, -) ( ).
, ,
. ,
.
.
, Ll ( ) ,
, .
. ,
- , .
. , , (
). :
,
. - (. 16).
48 :lO 5
.n .
( ) (
).
,
. ,
. ,
S - S- . ,
,
, ,
( )-.. . ,
~. -,
, ..
.
( );
«
» ,
.
,
,
(. 12), (. 23) (. 18).
. .
~ ~':";~
. ,
, . 5.8.
,
.. ~
,
(
), (, - . ), (Ag. , Hg) .
. . , (,
, ),
,
. . .
UR , Il
.
,
- , ,
.
. , . , .
(, )
; -
: 49
5.4. l1 - a-NH~, (' .1 ,.
-
Tyr Val
Ser Thr
11 , Uy R., Wold F.: Posttranslational covalent modification of proteins. Science: 1977: 198: 890, .
(, . 4.10). - . . 4. }l , . 3.3, (. 5.4 5.5).
; ,
.
, (.
5.9). - N- Gly, - - Asn; - N- - Ala .
- - . nPOll, -
5.5. lI lf.1 l( 8 1 '
-
Lys Lys
11 , Uy R., Wold F.: Posttranslational covalent modification of proteins. Sc;ence: 1977: 198: 890, .
• -. { __ ---L.~---7 _s_-s_J----I._2-0_21 6 I 11 I
s s I I s s
- { ---!L------1.L~- . 5.9. - - , ~
_1 .
, (. 51).
i; 124 , Lys N- Val -. , (. 5.1 ).
,
; . . , , , ,
. . -
48 . , .. .t 00 .. • 4 • ... " Jt ., . .. ' •
. 5.10. . .'1 lO .l
: . ~ . N-. ( Smyth . G .. Stein W. .• S.: The sequence of amino acid residues il1 bovinc pancreatic ribo nuclease. Revisions and conformaLions. J. 8;0/. Chem.
1963:238:227, .)
.
. ( ),
.
.
.
.
.
.
.
.
.
.
.
"l 5.6. ~~ ,"' ~:\1()DI
()
l05g .
ll
.
\1 ,
5 20%~ ...... 105g. ,
. ·
.
. . . .
. -
<l
IIrOToMepa
2 50000
- (--I,6-
I-~. 3.1.3.11) (L-:2- -
. 2.6.1.13) -- (-:2
[]. 6.4.1.3) () . (L-:
N- . 1.1.1.27) ( -,
3.6.1.3) . coli (L-: NH. []. 6.3.1.2) --Jl (- : 2 [],
6.4.1.2)
48500 4100000 409000
" Klotz 1. ., Langerman N.R., Darnall D. W.: Quaternary structure of enzymes. "". Rell • ioc/lcm. 1970:39:25.
21 .
( ). - - . .
5-15%. , , .
.
;
(.
~- ), ,
-
.
100000 .
, , .
. 5.6
, -
Advances in Protein Chemistry, Academic Press. 1944- 1987. (. )
Aisen .. Lis{m~'sky J. lron transport and storage proteins, l. Rev. Biochem .• 1980.49. 357.
Bald}~';n R. L. Intermcdiates in protein folding. . Rev. ih .. 1975, 44,453.
Chou . ., Fassman G. D. Empirical predictions of protein structure. . Rev. Biochem., 1978. 47, 251.
Cro}; L. R. Handbook of Amino Acid Sequences of Proteins. Joynson-Bruvvers Ltd. (Oxford. England), 1973.
Gurd F. N., Rothgeh . . Motions in proteins, Adv. Protein Chem., 1979, 33, 74. .
Hascllemeyer R. ., {leHarvell . Electron microscopy of enzy mes. . Rev. Biochem., 1974. 43. 279.
Lennar= w. J. (ed.) The Biochemistry ofGlycoproteins and Pro teoglycans, Plenum Press, 1980.
js ., Rossman . G. X-ray studies 01' protein interactions. . Rev. Biochem .. 1974. 43, 475.
Neuratll ., Hill R. L. (ed.) Thc Proteins, 3rd ed., Academic Press. 1975.
Osborne J. . Jr., ' . . Jr. Thc plasma lipoproteins, Adv. Protcin Chem., 1977. 31, 253.
Smitll L. ., ·n . J., GOf{O . . Jr. The plasma liporro teins: Structure and metabolism. . Rc\'. Biochcm .• 1978.47.751.
6
, ,
. ,
.
,
. , . , , ,
,
~- )
). ,
- reMo . , 2,3- ()
.
. -
,
, () (). (. 6.1), Q- . - -
~- . (), (V) () , : , V, , V, , Pr, Pr, (. 6.2). .' -
(Fe2+). : (Fe2+ Fe3+), , , , (Mg2+). ,
( ; . . 12). , Fe1+ .
. 6.1. . - . - ,
d, .
: ll 53
-
0-
. 6.2. . Ko.'Ibua , 't" , ; (Fe2+).
, . - :
CTOPl;my ( - ).
.
(,
) , ( ; . . 13).
Il
. 17000; 153 .
: , - Rp;
. , , (, Thr, Trp ., Tyr), , -
. ,
, ... Rp (, Leu, Val, Phe, Met).
,
,
4,5 3,5 2,5 (. 6.3). 75% -,
7 20 . N-, . , , , . , , , ,
QT N- . , His F8 - F, . ', (,
), ; , F8 () 7 () (. 6.3).
,
. 6.3. . - , . -: . ( Dickerson R. . In: The Proteins, 2nd ed., Vo). 2. Neurath . (editor). Academic Press, 1964, .)
54 6
;
; -
,
,
,
ll
3,5 (, ), - ,
.
. Fe2+
(
) . . , ,
,
. :
.
F; ,
,
His F8 His F7.
. His F8 (. 6.4). (His 7) ,
His ·F8. His 7 (. 6.4).
0,03 His F8.
,
0,01 . ,
, , His F8
n
His (7)
. 6.4. . .l , . ( . . et al .• Physiol6gische Chemie. Springcr-Verlag,
1975. .)
; .
Fe2+. d
.
.
1210 (. 6.5).
~O 0:;;.-'
I __ Fe
. 6.5. yr.lepoJ1L1. J1
( ).
: z 55
() 25000 . .
. ,
:
, 02? · , . , ,
, (Fe, , ) ,
(. 6.6). . (. 6.6). , ,
200 , - 02' (
1 %) .
IC
, ? ,
(<< »),
.
( , - ). -
1 ( ). (. 6.7).
[:5 [:5 ~O ~O
C~
J J
~) ~) . 6.6. . . 7 -
. 900 .
100
80
::r
20
100
, ,
J 100 . ., . 40 . ., 2- 20 . . 20 . .
, . ,
, 02
5 . .; ,
.
- - ,
· .. : 1) 02 ; 2) 02 Ica
.
,
.
, ,
,
( , ~,y, , S . .). .
6 "
- ~- - - 141 , ~--146; - ~ () .
~-, - -
.
~ , . ,
,
,
~- ,
' . - , , . ,
, (- )-
; -,
~-.
, , .
: (
)-2~2; HbF ( )-22; HbS ( )-2S2; 2 ( )-22 . ( ),
. ( . - , , ),
.
(
); , (. 6.8). , -
100
90
80
lf/. 70 60 s % CII 50 I :J 40
R u
::r
, . .
. 6.8. l .1() : ~.10. Jl .l0 :l :\1 1. .• - .10 40 :\1:\1 . ., ll - .10 20 . .; .1 , .1.1 Jll1 . ..., 5 . . 'J . l·0 II ll J . . (l0J1 .1 . :\1.10. ilHa.1or .1 .) ( Stanhury J. .. \Vyngaarden J. ., Fredrickson . S. (editors): The Metabolic Basis ol'lnherited Diseases. 4th ed. Gw-ilI. 1978, -
.)
02 Toro, 02' , 02 . ,
,
02 ~ 02 02'
. , ,
, = 100 . ., , 2 I
= 20 . . (. 6.8). 2 02 -
so - ' . so ,
2 - . (HBF). so = 26 . . ., HBF Pso = 20 . . F , .
HBF ; 02' .
: ~f ll 57
O~
, (. 6.9). 02'
. ,
. /-
/-, 02 (. 6.1 6.11).
feMor - ( . tut-);
(02) R- (re /d-) (. 6.12). R - ;
.
, ,
.
, 0,06 , (. 6.13).
, _ / I
I i
J I
Arg Asp Lys NH+ 0- .. ,
14'---1~ 40
~ . 6.9. . - . . . ( Stryer L.: Biochemistry, 2nd ed .•
Freeman. 1981. .)
,
COz . 02 ; 15% 02, , .
02 (. 6.14). - , .
1 / ) I ,
R -
. 6.10. - R- (2/2) 15r (u./P.). , . . (2/2) . u2/2-
u,/.- ( )
Tyr 7 42)
. 6.11. . . d ul,f32-KOH
. « ) , . . ( Perutz . F.: Molecular p<lthology fhu hemoglobin_ Stereochemical interpretation of l oxygen affinities_ Nalure 1971 :232:408, -
.)
___ -
08oro
. 6.13. , _ F8_ ( Stryer L: Biochemistry, 2nd ed., Free-
. 1981, .)
-
R-
. 6.12. - R- . .1 ;1 ( ), . . . . ( "lII). - R- <I"13 . . , . . IC. . . .1 - R- J1 , :. _ ( Perulz . F.: Hemoglobin strueture
and respiratory transporr. Sci. m_ [.] 1978:239:92, .)
: .'?1u ..lOUll 59
2 + 2 & • 2 ll lRR
llll - .
. (. 6.15). :
, .J -,
. ,
. , COZ• .
TeTpaMepHoro -bI , -
r'"'I"7"~~--.
. b.15. . , ll , , . . -- l>l ,·. l .:. -, 'J . } . ( ) -
.
.
, , ,
-; (146) ~-. ,
CO~ (. 6.15). l.
- ,
~ . . ( )
~-. R-
. . , ':'
.
(
).
. ,
. ,
-
-
60 6
J3-: ValNAl, LysEF6 His Hf 1 (. 6.17). . r - ror , e Il- - , . - R-.
or , ,
J3- 21 His, Ser, , . - - r or
. . R-
reMor r . ( 3000 /) .· , Fe2+
, .
His 21
. 6.17. . ~ P-u .. ( Amone .: X-ray diffrac110n study of bondl~g of 2.3- diphosphoglycera1e 10 human dhmgl. Nature
1972:237: 146, .)
, - J3-, .
( ), .
, .
, , l' .
ror
r or r - Il .
Fe3+ -, . , -
02' - R - - -. , . J3- ~ R- - , , .
,
R- ( ), , . .
, ( ).
S Glu Ai6)J3 Val. 2 (Glu Val)
, Glu l J3- « ». ,
S ( ). r r ror
, J3-, (. 6.18). S ,
\.:
. ror S .· .
: 61
S S
S
. 6.18. , S ( ) ( ) S. S . , '
. ( Stryer L.: Biochemistry, 2nd cd., Freeman, 1981, .)
S , .
. ,
S
S, or S «» . , - S. ( ), - R- . : S - ( S) , R-.
I-
. -.
S (. 6.18), «» S ,
. , r R- - S .
S .
(. 6.19).
. 6~19. ~~ S. ( Maugh . 11: new understanding of sickle 1l emerges.
Sc;ence 1981:211:265, .)
. 6.20. () 11 () lJ.. r , )( '\.1 ]I. '3. ,l ~ 8 (
), ~)l> I1f ~-6 II '~t ~II (.1. 36).
:
(. 6.20), .
-. -
(-) - (- ). ,
.
,
(. . 36).
Dean J .. Schechter . N. Sickle-cell anemia: ll and l ll basis of therapcutic approaches. (3 parts). N. Engl. J. Med., 1978, 299. 752. 804, 863.
Klot;: 1. .. n D. N.. inK L. . Ra1ional approaches 10 chemotherapy: Antisickling agents. Science. 1981,213. 724.
Perut;: . F. Hemog10gin structure and respiratory transport. Sci. m. (Dec.), 1978, 239, 92.
Perut;: . F. The rcgu1ation of oxygen-affinity of hemog10bin: I of structure of globin heme iron, . Rev. Biochem .• 1979, 48. 327.
Stamatoyannopoulo.'t G. The mll1l basis of hemoglobin di sease. . Rev. Genet., 1972. 6. 47.
Winslm~' R. . .4nderson /'. F. Thc hemoglobinopathies. Page 1666. In: The Metabolic Basis of Inhcrited Disease. 5th ed., Stanbury J. . et l. (eds.), Gw-iII. 1983.
7
;
,
aTc .
, . (+, -, )
. . ,
. .
l\tI
. ,
. (
)
(, ). , .
.
, - , . , ,
(). ~; , -
(), -; ,
(),- .
, ,
- .
. . . . · .
,
(IUB), .
. ,
; u .
. , {J IUB, .
1. ,
, ,
(
13). 2. :
- "f ( );
u
-.
3. Jl . , . , ,
L- + NADt = + 2 + NADH + t, 1.1.1.37 L-: NAD t ().
4. IIOMep
():
, -- .
. , 2.7.1.1 , 2 (). 7 ( ) 1 ( i ). , : D--6- , . . ,
64 7
.
. . .
1. . , - .
S80CC + S' I<CJ1 = SI<CJ1 + S' 8.
,
, -, -, = , NH2 -CH-NH-. :
1.1. , ll ( ). :
1.1.1.1. : NAD+ [ ]"
+ NAD+ = + NADH + +.
1.4. , CH-NH1
( ). : 1.4.1.3. L-: NAD(P)+
() [ ]. NAD(P)+ , NAD+ , NADP+.
L- + 2 + NAD(P)+ = = - + NH"'4 + NAD(P)H + +.
2. . , G ( ) S S':
S-G + S' = S' -G + S.
,
, ,
, ,
. : 2.3. QJ. : 2.3.1.6. -: -
[-]
2.7.1.1. : D- 6- [ ]
+ D- = + D--6-.·
3. . , m , , , , , - P-N. :.
3.1. , LI . :
3.1.1.8. - [ ]
+ 2 = + .
3.2.1.23. ~-- - [~ ]
~-D- + 2 = + D-.
, , , , N-. ,
. :
: , .
4. . , , .
I I - = - + =
, -, -, N, C-S -. :
4.1.2. -. : 4.1.2.7. -l-- []
-l- = + .
L- = + 2.
5. . ,
, .
.: 5.2. u-m-. : 5.2.1.3. -m- ll--m-
[]
-m- = ll--.
5.3.1.1. D--3---
[]
6. r. ( . -). . ,
.
. ,
-, C-S, C-N -. :
6.3. ,
]
6.4. ,
- ]
( ) ( ) . . « »
. ,
, - .
, (
IUB 1,2,5 6). , ,
, ( IUB 3 4).
, . . ,
,
, .
, -
(. 7.1).
-
3 1573
. 7.1. - , ro NADf- 8 ,
u--
.
:
NAOH NAO+. NAD+ ( , ) .
NAOH NAO+, . NAO+ . ,
. , , , , NAOH, (. 7.1).
m 7.1. ~ "Jii llltl NAD·
8
D-G + +:t A-G + . [ G , D-G, . ( ),
(
). :
D-GX A-G D CoE-G
( G), .
, :
D-HX COE
D CoE-
,
(. 1.1), D-HX - D CoE-
, _- :
NAD+, NADP+ FMN, FAD Q
-
- 6• B..L- lI, , Jl -
, ,
. .
. , ,
() (. . 34.1). NAD~ NADP+ .. 7.2.
« »
. « »
. , , , (, , «» , , ). . 1.3 ,
.
11
O---CH1
GJ'NH'
O=P-O-C~H I
. , , - . 7.1.. NAD(P)+; R = ( NAD+) , --2-( NADP+).
.wll: 67
. 7.3. « » H~ .
(
), , 1 3 .
, , ( ) . , 1 3, , -
. 1, 3 ( ). , ,
L-, , L-.
, , .
. ,
( , -
. .),
. , .
.
,
. .
~'1
. ,
-, L-. :
(, D- )
L-
.
, .
. : Il
, ( ~), .
II
, -
, - .
, ,l
- . .
, .
. ,
,
- , .
- .
NAD+, NADP+, . , , , ' (, ),
NADPH, , -
68 7
(, ), NAD+.
,
. ,
R
.
, ,
. npo
.
, .
. (, 10-6 ), (, 10-9 ) (, 10-12 ) ( ). (
.... U, nU pU.
" ;
, NAD+ NADP+ ( ),
, NADH NADPH ( NAD.f- NADP+) 340 (. 7.4).
NADH NAD+ ( ) (D) 340 , D (. 7.5).
(. 7.6)
( . 7.5) . . , J.I D 340 .
1,0
0,8 \ ..
8 I \ I \ ... I I 0,6 : I , :
11: I , .. : ,
. ... ,
. 7.4. NAD+ NADH. l1 44 /. I . NADP+ NADPH -
.
(NADH). (, . -
:::
~ ~ 0,85 :
.. ,
. 7.5. NADH- ND- . r 340 . . (S). .l0 340 . l- •• 10 NADH (J1 NADPH). 0.025- 0.2 .OJ- -
80 .
_wu: 69
0,05 0,10 0,15 0,20
... ,
. 7.6. _ !IO , _ 7.5, -
;I _
) ,
. -
. lll ,
(. 7.7).
~I
,
. ,
, , , .
, . lle
; - ~ .. - .
I( MeTO~
:
( ), (
, );
, - .
lll lll
. , .
, , (
)
, . lll .
lll 490- . 7.2.
. , ( 1 ) llM .
70 7
.
100000 g 98000 , 40---50%-
(NH4)2S04 90000 , 20---35%-
60000 80---11
- 58000 , 43-48%-
(NS. 52000 50000 wl 49000
, .
, , d . , ,
, .
~ ,
, .
,
.
, , ,
. , «» () 3-8 . , ,
. -
ll (. ).
~
, , % , Mr pU/Mr
10000 10 (100)
8000 12.2 98
1500 60 90
250 240 60
29 2000 58
20 2600 52 12 4160 50 10 4900 49
NAD+ .
, NAD ~ ,
, , « »,
, .
, Q (, ), , -
.
, <? , I< . .
(, ) .
.
, (NH4)2 S04~ u
.
. -
: ('n 71
( )
, . ( ')
I ( ) , . , ,
( ).
, (. . 2) . ,
, - .
.
,
(. . 2).
in situ (<<»). ( 2 10 ) , . , ,
. ;
.
.
«-6-», , ,
,
~
-6- Pj • ,
( , . / , . , ,
. ,
,
, . .
, .
«» (<<») ,
, , , ,
, . >, .
. , , , ,
. ,
.
, ,
~
.
.
8,6 ,
.
. .
- :
1) (, ); 2) (NAD+); 3) (,
);
5) ; ( ).
+ NAD+
72 . 7
L- •• ,,", """,,
PIIC. 7.8. , L-- - .
.
(. 7.8). 370, , \ (. 7.9).
(. 7.10). 1,.
' (, ) , .
. rr (. 130000) , (
. 34000). -
() SH - S ()
~~'~« NAD+ NADH + +
. 7.9. perpdMMC Jl
.
1. Is . , .
1, Is, 12' 1 14.
:
-
- - , -
(,
).
~
,
.
, ,
. ,
, , It. ,
. , ,
.
. ,
llm: ('ROlil"mflll 73
5 4 3 2 1
. 7.10. () . 8,6' , .
. . . . . . u . ( Dr. Melvin Black and Mr. Hugh Millcr, St Luke's
Hospital, San Francisco.)
.
. , ,- ,
, (
) . .
, ,
,
u , . . ,
.
8
.
. . 7.3 , .
, ,
,
,
.
. . ,
. , (Southern, 1975) -
74 7
il ,
(
;
, , .
. ,
(
; . . 6) , ,
, .
~- ~, (Dozy, Forman, l0, 1979; , Chang, Dozy, 1982). ,
, ~- ,
, ~ (Pirastu et l., 1984). u.- - ~ - .
u.- -
-, u.-
(Kidd et l., 1983).
, (. . 38). ,
(l), ~ (Val); ~- , 1 ~ Mst 11 Sau 1 (Orkin et l., 1982).
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~ . 36.
",: 75
ll II
. D .. Lardy ., Myrhiick . (eds.) The Enzymes, .. d ed., 7 vols. Academic P .. ess, 1970--1973.
Nm'd F. }'. (ed.) Advances in Enzymology, Interscience. [Is8ued anllually.]
Per.~/lt . Enzyme Stuctue and Mechanism, 2nd ed., Freeman, 1985.
Hirs . . W., imas('heff S. N. (eds.) Enzyme stucture. Pats - . In: MethodsjJ1 Enzymology, Vol. 11, 1967; Vols 25 d 26, 1972; Vol. 27. 1973; Vol. 47, 1977; Vols. 48 and 49, 1978: Vol. 49. 1979. Academic Press.
McCormick D. .. -'ig/lt L. D. (eds.) Vitamins d coenzymes, Parts A-F. l: Methods in Enzymology. Vol. 18, 1970; Vols 18 and 18, 1971; Vol. 62, 1979; Vols 66 and 67, 1980. Academic Press.
7.m Nmlt', 1978. Recommendations of the No menclature Committee the International Union of Bioc hcmisty I the NomenclatUI"e and Classification of Enzy mes. Academic Pess, 1979.
II
m .-. (ed.). Methods of Enzymatic Analysis, 2nd English ed. 4 vols, Academic Press, 1974.
. D., LardJ' ., Myrbiick . (eds.). Thc Enzymes. 3rd ed. 7 vols, Academic Press, 1970--1973.
l'ick S. ., Kaplan N. . (eds.). Methods in Enzymology, 69 vols, Academic Press, 1955·-1987.
.ffi'nn-Slnhff . ' al. Affinity hmtg .. h. Perga Press, 1978.
Jacoby W. . (ed.) Enzyme puification and related techniques. ': Methods in Enzymology, Vol. 22, Academic Pess, 1971.
Jacoby W. ., Wilchek . (eds.). Affinity techniques. In: Met hods in Enzymology, Vol. 34, 1974; Vol. 46, 1977, Acade mic Press.
Mosbach . (ed.). Immobilized enzymes. ': Methods in Enzy mology, Vol. 44, Academic Press. 1976.
De Pierre J. W., Ernster L. Enzyme topology of intracellular membranes, Annu. Rev. Biochem., 1977, 46, 201.
ll II
m . . Aspartate aminotransferase, Test of the Month 6, 1980, No. 2.
Bergstrom . Determination of serum alkaline phosphatase tivity, Test of the Month 1, 1974, No. 22.
Cavanee W. . ' al. Expression of recessive alleles chromo sl mechanisms in retinobIastomas, Natu .. e. 1983, 305, 779.
Dozy . ., Forman . N., Abuelo D. N. Penatal diagnosis of homozygous a-thalassemia, JAMA, 1979, 241, 1610.
Fishinger . F. Creatine phO!~phokinase and its izoenzymes, Test of the Month 2, 1976, No. 6.
Gusella J. F. , al. DNA markers for nervous system diseases, Science, 1984, 225, 1320.
. W., Chang J., Dozy . . Pages 275-283. ': Thalasse mia: RecencAdvances in Detection and Treatment, ., Carcassi U., Rowley . (eds.), . R. Liss, 1982.
Kidd V. J. , al. a,-Antitrypsin deficiency detection direct analysis of the mutation in the~gene, Nature, 1983, 304, 230.
Little . F. R. , al. Model for antenatal diagnosis of thalassemia and othe m~nogenic disoders molecular analysis linked DNA polymorphisms, Nature, 1980, 285, 144.
McNair R. D. Lactate dehydrogenase, Test of the Month 2, 1976. No. 3.
Orkin S. . ! al. Improved detection of the sickle mutation DNA analysis: Application to prenatal diagnosis, N. Engl. J. Med., 1982, 307, 32.
Pirastu . et al. Multiple mutations produce -thlssmi in Sardinia, Science, 1984, 223, 929.
Southern . . Detection of specific sequences among DNA fragments separated gel electrophoresis, J. Mol. Biol., 1975. 98. 503.
Wilk;nson J. . Clinical applications of isozymes, Clin. Chem., 1970. 16, 733.
Wilkinson J. . Clinical significance of enzyme activity measu rements, Clin. Chem.. 1970, 16, 882.
Woo S. L. . , al. Cloned human phenylalanine hydroxylase gene allows prenatal diagnosis and carrier detection of clas sical phenylketonuria, Nature, 1983, 306, 151.
8
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Christensen . N. Dissociation, Enzyrne Kinetics, Bioenergetics, Saunders, 1975.
Eng/e . . Enzyme Kinetics, Wiley. 1977. Piszki»'icz D. Kinetics of Chernical and Enzyrne-Catalyzed Re
aetions, Oxford Univ. Press, 1977. Segel 1. . Enzyrne Kinetics. Wiley, 1975. Sigman D .. S., Mooser G. Chernieal studies of ~rn active si
tes, . Rev. Biochern., 1975, 44, 889. Van Ta"'!len .. (ed.) Bioorganie Chernistry, Vol. 1, Enzyrne
AetlOn, 1977; Vol. 2, Maero- and Multirnoleeular Systerns, 1977; Vol. 3, Substrate Behavior. 1978, Academic Press.
9
. , n
, .
?
Robert . Murray, D, PhD Professor of Biochemistry University of Toronto
Daryl . Granner, MD Professor and Chairman Department of Molecular .Physiology and Biophysics Professor of Medicine Vanderbilt University Nashville, Tennessee
Peter A.Mayes, PhD, DSc Reader in Biochemistry Royal Veterinary College University of London
Vlctor W. Rodwell, PhD Professor of Biochemistry Purdue University West Lafayette, Indiana
APPLETON & LANGE Norwalk, Connecticut/San Mateo, California
.
.
.
.
1
. .-. . . . .
- . . .
«» 1993
577.2 20.070
: ., ., ., .
63 : 2- . . 1. . .: - .: , 1993. - 384 ., .
ISBN 5-03-001774-7
.
21-, - ,
.
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2- . :
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ISBN 5-03-001774-7 (.) ISBN 5-03-001773-9 ISBN 0-8385-3648-4 (r.)
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f.laBHbIx .l , .
Un.llu l'xu.uue .", 19
RR ( , ). 6.
. , ,
, ,
.
1930-x . , - . , , , 1 SN), -. ,
,
, ,
in vitro, ( , . .). , .
, - : ,
. «
).
,
. ,
, . 2.8. . in vivo in vitro . ,
JJ8. 2.8. ,
tlll
, .
, .
• , . ,
,
.
• - . . . -
( ).
. , ,
-+ -+ .
• . .
• . , . ;
, .
• .
, .
• .
• , .
• , , .
• .
• .
, ,
, ,
. ,
, , :
20 2
,
.
. -
.
, ,
. .
,
; , ,
. , , ~, ; ,
,
.
Freifelder D. Physical Biochemistry: Applications to Biochemi stry and Molecular Biology, Freeman, 1982.
Fruton J. S. Molecules and Life: Historical Essays the Inter play of Chemistry and Biology, Wiley-Interscience, 1972.
Weinberg R. . molecu1es oflife, Sci. m. (Oct.), 1985,253, 48. ( : .)
1
3
),
, , ,
.
'
- . L--R.
,
, ,
,
~x - cTpaHceM .
;
, . . 3.4 3.5.
ffi
, , . , -, ;
. ,
, ( ) (
).- , : ,
, [,
( )] .
.
;
.
.
- () (. 3.1).
300 , 20 . l 20 L-- (. 3.2). 20 - , . - ,
(. 30). ,
(. 3.4).
, R - , ,
- , .
1 -
.
-
( ). , , ( 7,0) , -
, L-r:
L--.
, -
-NH~. , , :
R-COOH +:t R-COO- + +, R-NH~~R-NH2+H+.
R - R - NH j J , .. , R-COO R-N2- (.. ) . R , R-N"- , R-COOH , R - NH j. , (7,4 7,1 ), , R-COO-. ()
, R - NH j. ,
, . 3.2,. , . 3.2., . , , , , . - - - 2 10 (. 3.1). , ' 2 • 99%. -
, R - NH~. , ,
(R-COO-). ,
, , . 3.2,.
u
. :
. =-log •.
. 3.1 20 , .
() ( ) . , .. J .
,
;
, . ,
,
(pl). , ,
, . 3.3 ..
I pI. , ,
I . , , pI . . (R-COOH) = 2,35 2 (R-Nj) = 9,69, (pI)
. + 2 2,35 + 9,69 pI = 2 = 2 = 6,02.
I , , ,
N +
. 3.2. . , . . , · ; .
- ~.
23
-
NH2 1 1:+
NH 1 11
(u) R-SH
, ,
. 3.4? , ,
, ,
lO ( uIO JI . 3.4). (), .. , ( . 3.4,). l-
.
}:- 11
. 3.3. -:)l: «»). , . l:l:
.
R-S- 8,3
pI = 2,09 i 3,86 = 2,98.
lO , . , ,
pI = ? + • 2
pI = 9,7, pI = 10,8. pI .
- , pI ,-
. lO lO .
, :-.
-
24 3
~ \\ 0-
} I = 29 NH + pKz - 3,86 ~s~ = 9,82 ~~ NH!
IHO/ \\ (- ) ~ • (IJ-) (-NH] .) - \\
-() \\
. 3.4. :I "" .
. , I 6 8 F 7,0, I = 6 , pI = 8 - .
, , , R-, ,- (.
3.2). . 3.3
, . (,
).
,
( ), (. 3.4 3.5). , , -
.1 3.2. JIc.tll L.,.u-n, , . <l .. 1 R-rpynn
(. 3.4) . ' 20 D-.. D- ,
; D- .
,
. (
, ) (, , ):
(> 2000 ). . , ,
.
,
, .. , .
BeRe
-
11
~KpaLeHHoe
Gly(G)
Ala(A)
Val(V)
Leu(L)
Ile(I)
cu ( ) Ser(S)
Thr(T)
Tyr(Y)
Cys(C)
Met(M)
. KUCJIbIe lC~ Asp(D)
Asn(N)
fl
--2 "
11 (l) (), lCfl . (. . 29 54). . flfl .
2) u. :
NH)f-
NH2
••
,..
.. . .
3.4. -, .
(2--4-
fH2-2-2-'- -.-
+N .N
+N I
(. 31)
,
(. 31)
(. 32)
- (3-)
(. 17)
(. 27)
- ( ) 2-2-1-
I ,
(. 32) +NH)
- (2--3-)
H)N+-CH~H-COO
(. 35) )
,
"'ma. = 570 . -,
, 1 . , ;
, 2• , 2 -. N ,
, -. ( 4-) .
(. 3.6) ,
. , , yr .
, , x . (. 3.7). , , -
. 3.6. .
YlYo-
. .
II, ,
, yr
.
29
R-bI , , R- , ;
- , R . R- "i
. , R ,
« »,
.
, -
,
= 7 .
(-SH) .
, :r .
-- , .
( , ).
, , ,
240 . . 3.8,
l
I1 ., ••
. 3.8. ii", -.
.
(cl. 3.6).
.
,
.
,
. , 5 . -· ,
(. 3.9).
, , , . -
l\ 3.6. \of .
lC
,
,
;
>.
. 3.9. ( - ).
, .
- .
( , ,
).
;
, ,
. ( ). ,
,
,
(
.5%-
90-J 100). (Leu, II. , Trp. Val, Met. Tyr) , (Pro, Ala, Gly) (Thr. Glu. Ser. Arg, Asp. His, Lys, Cys) (. 3.10). - . ,
(Gly, Ala, Val, Leu) ,
.
.
(
), Rr ( ). Rr ,
.
Rr, -
1
.
Rr, RA,a). .
.
, Rr•
() ;
(
) .
.
. , ,
900 (. 3.11).
. () -
.IIUm 31
2
J 11
. 3.11. , u (
Anal. Chem. 1953:25:396).
. ( ) ~.
,
.
. -
~
'1 :: 1-
s: ID s: :: 411 CJ ~ s s: U :5:
~ s: ~ 2 :: s: !. 1-
~ r:::: : ID > s: s ~ :: ct : '-s: s s: ...
~ L.
(
) , . , • .
,
. ,
. ,
N+-.
2 ,
Na +.
. . . ,
.
s: :5: ;s
:5: 1 ::
: :5: ;s
S
~
~ : 1- CJ
':5: : s: s: : ;s : 's
: :: 1- S > ;s :
'~ :r: ~ r::::
+--------.. ~I ,25)<11.I-------___ ~~ ... ---.... 4,25 .... 4f-----...
8. 3.12. ii - 50 ( 550) . .. 6 Jl (5 ~ 0,9 ); JI 5,28, 60 . . Jl ii . (55 0,9 ); Jl 3.25, ptl4,25. · . 6 . - 180 . , Jl 570 440 . Jl 440 , ( yt). ( . . Mertz, Purdue Uni-
versity.)
, yr (, )
.
. 0,5-2. 2000-5000 .
, , . . ,
, , . , .
. , , , ,
,- . «»
( , ) - ( .).
,
. 6,4 - 1 ;
. ,
, yr ,
, . , , 3,5 . .
Barrett G. . Chcrnistry and Biochernistry of the Arnino Acids, Chaprnan and , 1985.
. G. Tools· of Biochernistry, Wiley, 1977. Friedman . Chernistry and Biochernisrty of the Sulfhydryl
Group in Arnino Acids. Peptides, and Proteins, Pergamon Press, 1973.
Greenstein J. ., Winitz . Chernictry of the Arnino Acids, 3 vols, Wiley, 1961.
Heftman . Chrornatography: Laboratory Handbook of Chrornatographic and Electrophoretic Methods, 3rd ed., Van Nostrand, 1975. .
Touchstone J. . Practice of Thin Layer Chrornatography, Wi ley-Interscience, 1978.
Zweig G., Sherma J. Handbook of Chromatography, 2 vols, CRC Press, 1972.
4
. ,
. 10 n.
; .
. .
- . (, ) ( , ).
;
, ' ,
.
,
(;
, ,
.
. 4.1 , ,
. , n ,
.
2-1573
, C-OI ( )-. -
- .
, ,
, :- - .
(
) ,
, .
. 80- , «»
a-NH2- -- . . - . .
1. N- :
+ N 3
34 ' 4
- :
........... N 11
3. R- ( ) - -
:
. 4.2. .l 1 ;\- .l0 . <1 N-It fJ .... yTaMaT (Glu. ). - <1Jl
(Ala, ).
,
,
.
() 100 ,
. « »
,
, . 3.3 (. 4.2). .
, .
. - ,
(. 4.3).
Glu-Lys-(Ala, Gly, Tyr)-His-Ala
~
100
, (
; . . 6). .
, :
()
. 7,4 u . ,
- N-ubI
- (. J.I). .
, ,
, (
, ). - , - (.. t
- ). N- , , ( ' ),
, (. 4.1).
.'1u 4.1. · ;t.J ll .lII r-
() (N+)
nmu 35
(. .
). , ,
. ,
,,
(. 5). ,
(,
) (. 45, 48).
r
3 100 ), . , ,
, - 0-
L -, . ( )
,
, . .
. , , :
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
(. 4.4) ; , N
.
. , -
2*
SH
CHz - I
H--N + I -
«»
(. 5).
, , -, L , ,
. S -
, D- «IC) . .
(. 4.5). N- ,
.
11 I 11 Q ZJ
N C-N-<r--N
, ,
, ~
.
(. 3) ,
.
(30-100 ) .
. , ,
36 4
G L G QR L R N G D G N G N D G G N L S L
DLVAAEK
J.
,
. , ,
,
. , 1--4 (. 4.7).
. . 4.8
r , . 4.7. - ,
,
.
.
, . d-,
(2 = CH·CONH2) .
(),
. «»
(<<») ( 2-10%) ~ .
Ag+ (), () . .
.
-
(), ,
. -
;
.
. ·
. . .
.
nmu 37
~, t-
0.51-
. 4.7. - () . -50. IO/o- , 'l1 . - - . ( J. D. Pearson ! al., Department Biochemi-
stry, Purdue University.)
6 . HCl 110° . }_ .
. r .
, , . , q
(Val-Val, Il-Il, Val Il, Il-Val) 20 50%. , 24, 48, 72 96 .
.
, 96- . "Glx" ., Asx".
..
I 2~ ~CB-11 90 • I ' -2 1 oCb-
80 .. I ! ~ .. '-1 I
I 70 .. N Q. I N
l/l s 11: I 60 Q. S I .:
i .. u '- I s .. .. i
I 50
I N ~ 40 ~ :: ~
1
20
10
.
. 4.8. : ' () . - - . ( J. D. Pearson ct al., Department Biochemistry,
Purdue University.)
.
(. . 3.12) .
;
, , "
. -
- . , , -
38 4
. , 1- -2,4- (. 4.9), N- . ,
. ,
, .
,
,
(). 1967 .
N- <" . ,
, , , ,
.
~ . ,
CeHepa~ . ,
.
N01
t'HF
- S- .
N-ubl .
(.
38), .
,
. ,
;
'
. 8 . 38, - .
; , . .
(
, ).
.
(. 4.10). .
,
() , 20---60 .
. ,
, - ( 30 100 ). .
.
CNBr - .
nu 39
. 4.10. (J3TeHCHa). ;I ii (cleBa) ", l3- () . -
.
.
,
e .
. . . ,
( );
.
,
. CNBr .
. -. -
Trp-X. .
. . ' JIj Asn-Gly, .
. Staphylococcus aureus V8 Glu-, , - . Glu Lys . CNBr-.
. Jl . Asp"'Pro.
- , I!
Met, Trp, Arg Asn-Gly, ,
, Ji . - , < .
.
(. 4.7), (. 4.8) -.
r
( ); ·
N- ( ; . 4.11).
,
. . ,
, 30-40 ( 60 80 ) . ~ ' N lC n. iI,
N- i (. 4.11) np . .
. , ,
.
40 4
J.l.. ....... NHz
. 4.11. ( N- ).
. , ,
. , N- , -
JC JC .
,
.
,
(,
). ,
(. 4.12).
. 4.12. Z, JC
, , JC -, -.
m
, -, , .
, . ,
,
,
, .. .
1. , - ,
.
.
. ;
, .
4. 2 3 () .
, ,
.
5. .
. 3 . 8 - (21 ) 11 -- (30 ).
(124 ; . 5.10) 18 %; . -
nmu 41
, , ~ , , ,
, , . L- , ;
. ,
() (. 45).
Cantor R., Schimmel . R. Biophysical Chemistry, Part 1: The Conformation of Macromolecules, Freeman, 1980. [ : ., ' . .-: . 1984.]
Chin . ., Wold F. Separation of peptides phosph6cellu lose and other ceIlulose ion exchangers. Methods Enzymol, 1977, 47, 204.
, . G. The Tools of Biochemistry, Wiley, 1977. Craig L. , Cowburn D., Bleich . Methods of the study of
small polypeptide hormones and antibiotics in solution, . Rev. Biochem., 1975. 44, 509.
Dayhojj . (ed.) Atlas ofProtein Sequence and Structure, Vol. 5, National Biomedical Research Foundation, Washing ton, , 1972, Suppl. 1. 1973; Suppl. 2, 1976; Suppl. 3, 1979.
Hash J. . (ed.) Antibiotics. I: Methods in Enzymology, Vol. 43, Academic Press. 1975.
Heftman . Chromatography: Laboratory Handbook of
Chromatographic and Electrophoretic Methods, 3rd ed., Nostrand, 1975.
Jauregu;-Ade/l J., Mart; J. Acidic cleavage of the aspartyl prollne bond and the limitations of the reaction, Anal. Biochem., 1975,69,468.
Mahoney W. , Hermodson . . High-yield cleavage of tryp tophanyl peptide bonds o-iodosobenzoic acid, Bioche mistry, 1979, 18, 3810.
Mahoney W. , Smith . ; Hermodson . . Fragmentation of proteins with o-iodosobenzoic acid: Chemical mecha nism and identification of o-iodosobenzoic acid as reacti ve contaminant that modifies tyrosyl residues. Biochemi stry, 1981, 20. 443.
Marglin ., Merrifield R. . Chemical synthesis ofpeptides and proteins, . Rev. Biochem .. 1970, 39, 841.
Needelman S. . (ed.) Protein Sequence Determination, Sprin ger-Verlag, 1970.
Patthy L., Smith . L. Reversible modification of arginine res idues: Application to sequence studies restriction of tryptic hydrolysis to Iysine residues, J. Biol. Chem., 1975, 250,557.
Pearson J. D. et al. Reversed-phase supports for the resolution of large denatured protein fragments. J. Chromatogr., 1981, 207, 325.
Regnier . F., Gooding . High performance liquid chroma tography of proteins, Anal. Biochem .• 1980, 103. 1.
Snyder S. ., Innes R. . Peptide neurotransmitters. . Rev. Biochem., 1979. 48, 755.
Stewart J. ., Young J. D. Solid Phase Peptide Synthcsis. Free . 1969.
Storm D. R., Rosenthal S., Swanson . . Polymyxin antibio tics, . Rev. Biochem., 1977, 46, 723.
Zweig G., Sherma J. Handbook of Chromatography, 2 vols, CRC Press. 1972.
5
: ~
f
.
- . 8000-1 000. , -
: , ,
.
. ,
(. 6), (. 54) (. 26), . , , ,
(. 56),
( ) . .
,
. n
·
.
; 60 () , .
. , .
.
, _
, , , , .
, , 1907-1908 . . (. 5.1).
.l 5.1. ildl1 6C.1KOB. t'<l
:\10'-=
.
.
70-80%- ,
.
J.I
.
Gly. Ala. Pro
. , ,
. ,
, , ,
.
( ), . 1 3- 4. . ,
, . , 1 , ,·
. . ,
.
, (. 5.2). (),
,
(. 7).
, -. . l 5.2. tl
. .),
(), tl ( l), ()
. , .
; , 8,6 « » ,-, 2-, 13- -. ;
, , ,
(. 26). , .
,
, , , , F . , .
t , (.
). ,
,
. , , ,
«
» , ,
.
L-- . ,
, :
.
) ( , , . . ).
- ,
}'
(. 5.1). , .
44 5
/ .......... / ......... /'N/ N 1
~
. 5.1. ;
C-N.
5.1 ().
, , . . 5.2, , ,
,
.
, .
«»
( ),
. 5.2. . , , . -, -R- . , - - , ( ). , - , · ( , ) . - 0,36 . " ( ). ( , ulig L., Corey R. , Branson . R.: The structure of proteins. Two hydrogen-bonded helical figur tions of the polypeptide chain. Proc. Natl. Acad. Sc;. USA
.1951:37:205.)
. , . ( S-S) J3- ( S-S ) , ;
(. . 4.10).
tl
1) , ; 2) ,
; 3) ,
, .
, . .
(. . «u-) «
J3-»).
.
, . · .
,
. , + 1, - 1. , , .
(, + -) , ,
.
"
,
, .
.
, 1930- ., - 0,5--
0--
. 5.3. " -.
O,5r
. 5.4. - . R - , . ", , " . ( Stryer L. Biochemistry, 2nd ed., Freemal1, 1981. : . I:). 3- .- : . 1985.)
0,55 . , .
(. 5.2). m , , -
- (. 5.3). R- -
(. 5.4). 3,6 , 0,54 , 0,5-{),55 , . , , 0,15 , . '(f (. 5.5).
1. - ,
, ,
.
2. . -.
3. ,
" ( ).
4. - ,
.
5. L- , , - .
46 5
R
. 5.5. -. - ~ R~ ~ ~ ( ). ( Haggis . . et l., Introduetion to
Molccul~r Biology. Wilcy. 1964.)
-,
- . (
,
N - ), R-,
- (. 5.3).
-
- - ( , -).
-
, - (. 5.6). ,
5.3. - -
Cys l
Val
(
N- ), LI
- ( . 5.6). ,
- ( ).
---
-
--- -
. 5.6. .l -. J'. ,10 NH- . (R) . clTOMbI , - , -
. ( Stryer L. Biochemistry. 2nd cd .• Free- , 1981. .)
: 47
. S.7. .n - /l. ~ Jl 124 . . -u- , . . (. 8) POJ-4• ( Kartha G., J., Harker .: Tertiary structure ofribo nuclcase. Nature 1967:213:862.) -
.
. . 5.7 ,
~-CTpYKTypa . . -, - (. 5.7).
-
, ' ,
~-CTpYKTypa ,
. . 5.7.
()
, ,
. . 5.7, . « » :
,
. , - ,
- -.
, (. 4). (
) , .
.
(-, -) ( ).
, ,
. ,
.
.
, Ll ( ) ,
, .
. ,
- , .
. , , (
). :
,
. - (. 16).
48 :lO 5
.n .
( ) (
).
,
. ,
. ,
S - S- . ,
,
, ,
( )-.. . ,
~. -,
, ..
.
( );
«
» ,
.
,
,
(. 12), (. 23) (. 18).
. .
~ ~':";~
. ,
, . 5.8.
,
.. ~
,
(
), (, - . ), (Ag. , Hg) .
. . , (,
, ),
,
. . .
UR , Il
.
,
- , ,
.
. , . , .
(, )
; -
: 49
5.4. l1 - a-NH~, (' .1 ,.
-
Tyr Val
Ser Thr
11 , Uy R., Wold F.: Posttranslational covalent modification of proteins. Science: 1977: 198: 890, .
(, . 4.10). - . . 4. }l , . 3.3, (. 5.4 5.5).
; ,
.
, (.
5.9). - N- Gly, - - Asn; - N- - Ala .
- - . nPOll, -
5.5. lI lf.1 l( 8 1 '
-
Lys Lys
11 , Uy R., Wold F.: Posttranslational covalent modification of proteins. Sc;ence: 1977: 198: 890, .
• -. { __ ---L.~---7 _s_-s_J----I._2-0_21 6 I 11 I
s s I I s s
- { ---!L------1.L~- . 5.9. - - , ~
_1 .
, (. 51).
i; 124 , Lys N- Val -. , (. 5.1 ).
,
; . . , , , ,
. . -
48 . , .. .t 00 .. • 4 • ... " Jt ., . .. ' •
. 5.10. . .'1 lO .l
: . ~ . N-. ( Smyth . G .. Stein W. .• S.: The sequence of amino acid residues il1 bovinc pancreatic ribo nuclease. Revisions and conformaLions. J. 8;0/. Chem.
1963:238:227, .)
.
. ( ),
.
.
.
.
.
.
.
.
.
.
.
"l 5.6. ~~ ,"' ~:\1()DI
()
l05g .
ll
.
\1 ,
5 20%~ ...... 105g. ,
. ·
.
. . . .
. -
<l
IIrOToMepa
2 50000
- (--I,6-
I-~. 3.1.3.11) (L-:2- -
. 2.6.1.13) -- (-:2
[]. 6.4.1.3) () . (L-:
N- . 1.1.1.27) ( -,
3.6.1.3) . coli (L-: NH. []. 6.3.1.2) --Jl (- : 2 [],
6.4.1.2)
48500 4100000 409000
" Klotz 1. ., Langerman N.R., Darnall D. W.: Quaternary structure of enzymes. "". Rell • ioc/lcm. 1970:39:25.
21 .
( ). - - . .
5-15%. , , .
.
;
(.
~- ), ,
-
.
100000 .
, , .
. 5.6
, -
Advances in Protein Chemistry, Academic Press. 1944- 1987. (. )
Aisen .. Lis{m~'sky J. lron transport and storage proteins, l. Rev. Biochem .• 1980.49. 357.
Bald}~';n R. L. Intermcdiates in protein folding. . Rev. ih .. 1975, 44,453.
Chou . ., Fassman G. D. Empirical predictions of protein structure. . Rev. Biochem., 1978. 47, 251.
Cro}; L. R. Handbook of Amino Acid Sequences of Proteins. Joynson-Bruvvers Ltd. (Oxford. England), 1973.
Gurd F. N., Rothgeh . . Motions in proteins, Adv. Protein Chem., 1979, 33, 74. .
Hascllemeyer R. ., {leHarvell . Electron microscopy of enzy mes. . Rev. Biochem., 1974. 43. 279.
Lennar= w. J. (ed.) The Biochemistry ofGlycoproteins and Pro teoglycans, Plenum Press, 1980.
js ., Rossman . G. X-ray studies 01' protein interactions. . Rev. Biochem .. 1974. 43, 475.
Neuratll ., Hill R. L. (ed.) Thc Proteins, 3rd ed., Academic Press. 1975.
Osborne J. . Jr., ' . . Jr. Thc plasma lipoproteins, Adv. Protcin Chem., 1977. 31, 253.
Smitll L. ., ·n . J., GOf{O . . Jr. The plasma liporro teins: Structure and metabolism. . Rc\'. Biochcm .• 1978.47.751.
6
, ,
. ,
.
,
. , . , , ,
,
~- )
). ,
- reMo . , 2,3- ()
.
. -
,
, () (). (. 6.1), Q- . - -
~- . (), (V) () , : , V, , V, , Pr, Pr, (. 6.2). .' -
(Fe2+). : (Fe2+ Fe3+), , , , (Mg2+). ,
( ; . . 12). , Fe1+ .
. 6.1. . - . - ,
d, .
: ll 53
-
0-
. 6.2. . Ko.'Ibua , 't" , ; (Fe2+).
, . - :
CTOPl;my ( - ).
.
(,
) , ( ; . . 13).
Il
. 17000; 153 .
: , - Rp;
. , , (, Thr, Trp ., Tyr), , -
. ,
, ... Rp (, Leu, Val, Phe, Met).
,
,
4,5 3,5 2,5 (. 6.3). 75% -,
7 20 . N-, . , , , . , , , ,
QT N- . , His F8 - F, . ', (,
), ; , F8 () 7 () (. 6.3).
,
. 6.3. . - , . -: . ( Dickerson R. . In: The Proteins, 2nd ed., Vo). 2. Neurath . (editor). Academic Press, 1964, .)
54 6
;
; -
,
,
,
ll
3,5 (, ), - ,
.
. Fe2+
(
) . . , ,
,
. :
.
F; ,
,
His F8 His F7.
. His F8 (. 6.4). (His 7) ,
His ·F8. His 7 (. 6.4).
0,03 His F8.
,
0,01 . ,
, , His F8
n
His (7)
. 6.4. . .l , . ( . . et al .• Physiol6gische Chemie. Springcr-Verlag,
1975. .)
; .
Fe2+. d
.
.
1210 (. 6.5).
~O 0:;;.-'
I __ Fe
. 6.5. yr.lepoJ1L1. J1
( ).
: z 55
() 25000 . .
. ,
:
, 02? · , . , ,
, (Fe, , ) ,
(. 6.6). . (. 6.6). , ,
200 , - 02' (
1 %) .
IC
, ? ,
(<< »),
.
( , - ). -
1 ( ). (. 6.7).
[:5 [:5 ~O ~O
C~
J J
~) ~) . 6.6. . . 7 -
. 900 .
100
80
::r
20
100
, ,
J 100 . ., . 40 . ., 2- 20 . . 20 . .
, . ,
, 02
5 . .; ,
.
- - ,
· .. : 1) 02 ; 2) 02 Ica
.
,
.
, ,
,
( , ~,y, , S . .). .
6 "
- ~- - - 141 , ~--146; - ~ () .
~-, - -
.
~ , . ,
,
,
~- ,
' . - , , . ,
, (- )-
; -,
~-.
, , .
: (
)-2~2; HbF ( )-22; HbS ( )-2S2; 2 ( )-22 . ( ),
. ( . - , , ),
.
(
); , (. 6.8). , -
100
90
80
lf/. 70 60 s % CII 50 I :J 40
R u
::r
, . .
. 6.8. l .1() : ~.10. Jl .l0 :l :\1 1. .• - .10 40 :\1:\1 . ., ll - .10 20 . .; .1 , .1.1 Jll1 . ..., 5 . . 'J . l·0 II ll J . . (l0J1 .1 . :\1.10. ilHa.1or .1 .) ( Stanhury J. .. \Vyngaarden J. ., Fredrickson . S. (editors): The Metabolic Basis ol'lnherited Diseases. 4th ed. Gw-ilI. 1978, -
.)
02 Toro, 02' , 02 . ,
,
02 ~ 02 02'
. , ,
, = 100 . ., , 2 I
= 20 . . (. 6.8). 2 02 -
so - ' . so ,
2 - . (HBF). so = 26 . . ., HBF Pso = 20 . . F , .
HBF ; 02' .
: ~f ll 57
O~
, (. 6.9). 02'
. ,
. /-
/-, 02 (. 6.1 6.11).
feMor - ( . tut-);
(02) R- (re /d-) (. 6.12). R - ;
.
, ,
.
, 0,06 , (. 6.13).
, _ / I
I i
J I
Arg Asp Lys NH+ 0- .. ,
14'---1~ 40
~ . 6.9. . - . . . ( Stryer L.: Biochemistry, 2nd ed .•
Freeman. 1981. .)
,
COz . 02 ; 15% 02, , .
02 (. 6.14). - , .
1 / ) I ,
R -
. 6.10. - R- (2/2) 15r (u./P.). , . . (2/2) . u2/2-
u,/.- ( )
Tyr 7 42)
. 6.11. . . d ul,f32-KOH
. « ) , . . ( Perutz . F.: Molecular p<lthology fhu hemoglobin_ Stereochemical interpretation of l oxygen affinities_ Nalure 1971 :232:408, -
.)
___ -
08oro
. 6.13. , _ F8_ ( Stryer L: Biochemistry, 2nd ed., Free-
. 1981, .)
-
R-
. 6.12. - R- . .1 ;1 ( ), . . . . ( "lII). - R- <I"13 . . , . . IC. . . .1 - R- J1 , :. _ ( Perulz . F.: Hemoglobin strueture
and respiratory transporr. Sci. m_ [.] 1978:239:92, .)
: .'?1u ..lOUll 59
2 + 2 & • 2 ll lRR
llll - .
. (. 6.15). :
, .J -,
. ,
. , COZ• .
TeTpaMepHoro -bI , -
r'"'I"7"~~--.
. b.15. . , ll , , . . -- l>l ,·. l .:. -, 'J . } . ( ) -
.
.
, , ,
-; (146) ~-. ,
CO~ (. 6.15). l.
- ,
~ . . ( )
~-. R-
. . , ':'
.
(
).
. ,
. ,
-
-
60 6
J3-: ValNAl, LysEF6 His Hf 1 (. 6.17). . r - ror , e Il- - , . - R-.
or , ,
J3- 21 His, Ser, , . - - r or
. . R-
reMor r . ( 3000 /) .· , Fe2+
, .
His 21
. 6.17. . ~ P-u .. ( Amone .: X-ray diffrac110n study of bondl~g of 2.3- diphosphoglycera1e 10 human dhmgl. Nature
1972:237: 146, .)
, - J3-, .
( ), .
, .
, , l' .
ror
r or r - Il .
Fe3+ -, . , -
02' - R - - -. , . J3- ~ R- - , , .
,
R- ( ), , . .
, ( ).
S Glu Ai6)J3 Val. 2 (Glu Val)
, Glu l J3- « ». ,
S ( ). r r ror
, J3-, (. 6.18). S ,
\.:
. ror S .· .
: 61
S S
S
. 6.18. , S ( ) ( ) S. S . , '
. ( Stryer L.: Biochemistry, 2nd cd., Freeman, 1981, .)
S , .
. ,
S
S, or S «» . , - S. ( ), - R- . : S - ( S) , R-.
I-
. -.
S (. 6.18), «» S ,
. , r R- - S .
S .
(. 6.19).
. 6~19. ~~ S. ( Maugh . 11: new understanding of sickle 1l emerges.
Sc;ence 1981:211:265, .)
. 6.20. () 11 () lJ.. r , )( '\.1 ]I. '3. ,l ~ 8 (
), ~)l> I1f ~-6 II '~t ~II (.1. 36).
:
(. 6.20), .
-. -
(-) - (- ). ,
.
,
(. . 36).
Dean J .. Schechter . N. Sickle-cell anemia: ll and l ll basis of therapcutic approaches. (3 parts). N. Engl. J. Med., 1978, 299. 752. 804, 863.
Klot;: 1. .. n D. N.. inK L. . Ra1ional approaches 10 chemotherapy: Antisickling agents. Science. 1981,213. 724.
Perut;: . F. Hemog10gin structure and respiratory transport. Sci. m. (Dec.), 1978, 239, 92.
Perut;: . F. The rcgu1ation of oxygen-affinity of hemog10bin: I of structure of globin heme iron, . Rev. Biochem .• 1979, 48. 327.
Stamatoyannopoulo.'t G. The mll1l basis of hemoglobin di sease. . Rev. Genet., 1972. 6. 47.
Winslm~' R. . .4nderson /'. F. Thc hemoglobinopathies. Page 1666. In: The Metabolic Basis of Inhcrited Disease. 5th ed., Stanbury J. . et l. (eds.), Gw-iII. 1983.
7
;
,
aTc .
, . (+, -, )
. . ,
. .
l\tI
. ,
. (
)
(, ). , .
.
, - , . , ,
(). ~; , -
(), -; ,
(),- .
, ,
- .
. . . . · .
,
(IUB), .
. ,
; u .
. , {J IUB, .
1. ,
, ,
(
13). 2. :
- "f ( );
u
-.
3. Jl . , . , ,
L- + NADt = + 2 + NADH + t, 1.1.1.37 L-: NAD t ().
4. IIOMep
():
, -- .
. , 2.7.1.1 , 2 (). 7 ( ) 1 ( i ). , : D--6- , . . ,
64 7
.
. . .
1. . , - .
S80CC + S' I<CJ1 = SI<CJ1 + S' 8.
,
, -, -, = , NH2 -CH-NH-. :
1.1. , ll ( ). :
1.1.1.1. : NAD+ [ ]"
+ NAD+ = + NADH + +.
1.4. , CH-NH1
( ). : 1.4.1.3. L-: NAD(P)+
() [ ]. NAD(P)+ , NAD+ , NADP+.
L- + 2 + NAD(P)+ = = - + NH"'4 + NAD(P)H + +.
2. . , G ( ) S S':
S-G + S' = S' -G + S.
,
, ,
, ,
. : 2.3. QJ. : 2.3.1.6. -: -
[-]
2.7.1.1. : D- 6- [ ]
+ D- = + D--6-.·
3. . , m , , , , , - P-N. :.
3.1. , LI . :
3.1.1.8. - [ ]
+ 2 = + .
3.2.1.23. ~-- - [~ ]
~-D- + 2 = + D-.
, , , , N-. ,
. :
: , .
4. . , , .
I I - = - + =
, -, -, N, C-S -. :
4.1.2. -. : 4.1.2.7. -l-- []
-l- = + .
L- = + 2.
5. . ,
, .
.: 5.2. u-m-. : 5.2.1.3. -m- ll--m-
[]
-m- = ll--.
5.3.1.1. D--3---
[]
6. r. ( . -). . ,
.
. ,
-, C-S, C-N -. :
6.3. ,
]
6.4. ,
- ]
( ) ( ) . . « »
. ,
, - .
, (
IUB 1,2,5 6). , ,
, ( IUB 3 4).
, . . ,
,
, .
, -
(. 7.1).
-
3 1573
. 7.1. - , ro NADf- 8 ,
u--
.
:
NAOH NAO+. NAD+ ( , ) .
NAOH NAO+, . NAO+ . ,
. , , , , NAOH, (. 7.1).
m 7.1. ~ "Jii llltl NAD·
8
D-G + +:t A-G + . [ G , D-G, . ( ),
(
). :
D-GX A-G D CoE-G
( G), .
, :
D-HX COE
D CoE-
,
(. 1.1), D-HX - D CoE-
, _- :
NAD+, NADP+ FMN, FAD Q
-
- 6• B..L- lI, , Jl -
, ,
. .
. , ,
() (. . 34.1). NAD~ NADP+ .. 7.2.
« »
. « »
. , , , (, , «» , , ). . 1.3 ,
.
11
O---CH1
GJ'NH'
O=P-O-C~H I
. , , - . 7.1.. NAD(P)+; R = ( NAD+) , --2-( NADP+).
.wll: 67
. 7.3. « » H~ .
(
), , 1 3 .
, , ( ) . , 1 3, , -
. 1, 3 ( ). , ,
L-, , L-.
, , .
. ,
( , -
. .),
. , .
.
,
. .
~'1
. ,
-, L-. :
(, D- )
L-
.
, .
. : Il
, ( ~), .
II
, -
, - .
, ,l
- . .
, .
. ,
,
- , .
- .
NAD+, NADP+, . , , , ' (, ),
NADPH, , -
68 7
(, ), NAD+.
,
. ,
R
.
, ,
. npo
.
, .
. (, 10-6 ), (, 10-9 ) (, 10-12 ) ( ). (
.... U, nU pU.
" ;
, NAD+ NADP+ ( ),
, NADH NADPH ( NAD.f- NADP+) 340 (. 7.4).
NADH NAD+ ( ) (D) 340 , D (. 7.5).
(. 7.6)
( . 7.5) . . , J.I D 340 .
1,0
0,8 \ ..
8 I \ I \ ... I I 0,6 : I , :
11: I , .. : ,
. ... ,
. 7.4. NAD+ NADH. l1 44 /. I . NADP+ NADPH -
.
(NADH). (, . -
:::
~ ~ 0,85 :
.. ,
. 7.5. NADH- ND- . r 340 . . (S). .l0 340 . l- •• 10 NADH (J1 NADPH). 0.025- 0.2 .OJ- -
80 .
_wu: 69
0,05 0,10 0,15 0,20
... ,
. 7.6. _ !IO , _ 7.5, -
;I _
) ,
. -
. lll ,
(. 7.7).
~I
,
. ,
, , , .
, . lle
; - ~ .. - .
I( MeTO~
:
( ), (
, );
, - .
lll lll
. , .
, , (
)
, . lll .
lll 490- . 7.2.
. , ( 1 ) llM .
70 7
.
100000 g 98000 , 40---50%-
(NH4)2S04 90000 , 20---35%-
60000 80---11
- 58000 , 43-48%-
(NS. 52000 50000 wl 49000
, .
, , d . , ,
, .
~ ,
, .
,
.
, , ,
. , «» () 3-8 . , ,
. -
ll (. ).
~
, , % , Mr pU/Mr
10000 10 (100)
8000 12.2 98
1500 60 90
250 240 60
29 2000 58
20 2600 52 12 4160 50 10 4900 49
NAD+ .
, NAD ~ ,
, , « »,
, .
, Q (, ), , -
.
, <? , I< . .
(, ) .
.
, (NH4)2 S04~ u
.
. -
: ('n 71
( )
, . ( ')
I ( ) , . , ,
( ).
, (. . 2) . ,
, - .
.
,
(. . 2).
in situ (<<»). ( 2 10 ) , . , ,
. ;
.
.
«-6-», , ,
,
~
-6- Pj • ,
( , . / , . , ,
. ,
,
, . .
, .
«» (<<») ,
, , , ,
, . >, .
. , , , ,
. ,
.
, ,
~
.
.
8,6 ,
.
. .
- :
1) (, ); 2) (NAD+); 3) (,
);
5) ; ( ).
+ NAD+
72 . 7
L- •• ,,", """,,
PIIC. 7.8. , L-- - .
.
(. 7.8). 370, , \ (. 7.9).
(. 7.10). 1,.
' (, ) , .
. rr (. 130000) , (
. 34000). -
() SH - S ()
~~'~« NAD+ NADH + +
. 7.9. perpdMMC Jl
.
1. Is . , .
1, Is, 12' 1 14.
:
-
- - , -
(,
).
~
,
.
, ,
. ,
, , It. ,
. , ,
.
. ,
llm: ('ROlil"mflll 73
5 4 3 2 1
. 7.10. () . 8,6' , .
. . . . . . u . ( Dr. Melvin Black and Mr. Hugh Millcr, St Luke's
Hospital, San Francisco.)
.
. , ,- ,
, (
) . .
, ,
,
u , . . ,
.
8
.
. . 7.3 , .
, ,
,
,
.
. . ,
. , (Southern, 1975) -
74 7
il ,
(
;
, , .
. ,
(
; . . 6) , ,
, .
~- ~, (Dozy, Forman, l0, 1979; , Chang, Dozy, 1982). ,
, ~- ,
, ~ (Pirastu et l., 1984). u.- - ~ - .
u.- -
-, u.-
(Kidd et l., 1983).
, (. . 38). ,
(l), ~ (Val); ~- , 1 ~ Mst 11 Sau 1 (Orkin et l., 1982).
- , ,
.
(
).
( ),
. (). ( , ). , ,
, .
( 1), ~- ( Hind 111 1) (Little et l .• 1980; Woo et l., 1983).
, , (Woo et l., 1983) (. . 31). , , . , , .
,
He' .
, (Cavenee et l., 1983) (GuseIla et l., 1984).
~ . 36.
",: 75
ll II
. D .. Lardy ., Myrhiick . (eds.) The Enzymes, .. d ed., 7 vols. Academic P .. ess, 1970--1973.
Nm'd F. }'. (ed.) Advances in Enzymology, Interscience. [Is8ued anllually.]
Per.~/lt . Enzyme Stuctue and Mechanism, 2nd ed., Freeman, 1985.
Hirs . . W., imas('heff S. N. (eds.) Enzyme stucture. Pats - . In: MethodsjJ1 Enzymology, Vol. 11, 1967; Vols 25 d 26, 1972; Vol. 27. 1973; Vol. 47, 1977; Vols. 48 and 49, 1978: Vol. 49. 1979. Academic Press.
McCormick D. .. -'ig/lt L. D. (eds.) Vitamins d coenzymes, Parts A-F. l: Methods in Enzymology. Vol. 18, 1970; Vols 18 and 18, 1971; Vol. 62, 1979; Vols 66 and 67, 1980. Academic Press.
7.m Nmlt', 1978. Recommendations of the No menclature Committee the International Union of Bioc hcmisty I the NomenclatUI"e and Classification of Enzy mes. Academic Pess, 1979.
II
m .-. (ed.). Methods of Enzymatic Analysis, 2nd English ed. 4 vols, Academic Press, 1974.
. D., LardJ' ., Myrbiick . (eds.). Thc Enzymes. 3rd ed. 7 vols, Academic Press, 1970--1973.
l'ick S. ., Kaplan N. . (eds.). Methods in Enzymology, 69 vols, Academic Press, 1955·-1987.
.ffi'nn-Slnhff . ' al. Affinity hmtg .. h. Perga Press, 1978.
Jacoby W. . (ed.) Enzyme puification and related techniques. ': Methods in Enzymology, Vol. 22, Academic Pess, 1971.
Jacoby W. ., Wilchek . (eds.). Affinity techniques. In: Met hods in Enzymology, Vol. 34, 1974; Vol. 46, 1977, Acade mic Press.
Mosbach . (ed.). Immobilized enzymes. ': Methods in Enzy mology, Vol. 44, Academic Press. 1976.
De Pierre J. W., Ernster L. Enzyme topology of intracellular membranes, Annu. Rev. Biochem., 1977, 46, 201.
ll II
m . . Aspartate aminotransferase, Test of the Month 6, 1980, No. 2.
Bergstrom . Determination of serum alkaline phosphatase tivity, Test of the Month 1, 1974, No. 22.
Cavanee W. . ' al. Expression of recessive alleles chromo sl mechanisms in retinobIastomas, Natu .. e. 1983, 305, 779.
Dozy . ., Forman . N., Abuelo D. N. Penatal diagnosis of homozygous a-thalassemia, JAMA, 1979, 241, 1610.
Fishinger . F. Creatine phO!~phokinase and its izoenzymes, Test of the Month 2, 1976, No. 6.
Gusella J. F. , al. DNA markers for nervous system diseases, Science, 1984, 225, 1320.
. W., Chang J., Dozy . . Pages 275-283. ': Thalasse mia: RecencAdvances in Detection and Treatment, ., Carcassi U., Rowley . (eds.), . R. Liss, 1982.
Kidd V. J. , al. a,-Antitrypsin deficiency detection direct analysis of the mutation in the~gene, Nature, 1983, 304, 230.
Little . F. R. , al. Model for antenatal diagnosis of thalassemia and othe m~nogenic disoders molecular analysis linked DNA polymorphisms, Nature, 1980, 285, 144.
McNair R. D. Lactate dehydrogenase, Test of the Month 2, 1976. No. 3.
Orkin S. . ! al. Improved detection of the sickle mutation DNA analysis: Application to prenatal diagnosis, N. Engl. J. Med., 1982, 307, 32.
Pirastu . et al. Multiple mutations produce -thlssmi in Sardinia, Science, 1984, 223, 929.
Southern . . Detection of specific sequences among DNA fragments separated gel electrophoresis, J. Mol. Biol., 1975. 98. 503.
Wilk;nson J. . Clinical applications of isozymes, Clin. Chem., 1970. 16, 733.
Wilkinson J. . Clinical significance of enzyme activity measu rements, Clin. Chem.. 1970, 16, 882.
Woo S. L. . , al. Cloned human phenylalanine hydroxylase gene allows prenatal diagnosis and carrier detection of clas sical phenylketonuria, Nature, 1983, 306, 151.
8
. . 5 11 , , .
- , .
, - , , ,-
. ,
. , . , (
)-, ,
- , (~,
opaHO~
). -
1 . . ,
.
. ,
-
, .
Ix
-r ,
:
Y+R-X~Y-R+X.
u: 1) COCTOJIJI, R , ; 2) :
+ R - XpY···R···XpY - R + . '--Y--J
, ,
. I!GF , , I!GD ,
:
[Y][R-X]
I!G o
, I!G, :
I!G = I1GF + AGD•
, , AG I!GF
I1GD • , , I1G. , .
.\f(!llm: 1\ullt'm""'a 77
I!GF I!Go, , (I!G) (. . ). .
,
. 8.1 8.2; « », I!G, 6GF I!Go. . I1G . 8.1 (I!G < ). . 8.2 (I!G> ); I!GF (I!GF > ), AGD
(I!G D < ). , , I!G I!GF I!Go'
. (. 8.3).
. , -
G<
. (L\ G < ).
. 8.2. II .
" (6 G> ).
. 8.3. l, ;l . ['" R'" ];1 [ ... R'" X]II; L\ (;:' L\ G~-u
1 UJ1.
[Y"·R· .. X]b, , , [Y"·R"·X]B. ['" R .. · ] , Tor ['" R ... ]_ .
, , I!Gp , I!G : .
:
I!CO = - RT ln Keq.
, yr JJR .
, . .
. 1. r r, .. , .
2. (.. ),
r. .
, . , . , ,
. .
, ,
,
,
.
. 8.4. , -, -
, .
78 8
' ,
, .
. ,
, .
(. . ) . , (. . ), AG < , . ,
, .
x
; , , . . , ,
. 7:
D - G + A~A - G + , G , D - ,
. D - , -, - .
, :
D-GZ ~ A-G
. 1.
, _ . 2. ,
D - d . 3. -
(. .
),
(. .
1: 1).
,
, , . , (, -6- l-) , D :
S-y Enz '
- t
nz - S J'I . · .
:
D:G~E~~:G
Enz-G Enz-G··
\. Z-G·J Enz - , Enz - G * Enz - ** -- Enz - S, .
, u . , ( ) . . . .
. . :J
. ,
. :
+ +:t
.
: llmu 79
[R][C] Kd =----
. R - , Kd •
:
.
,
.
,
,
(Kd ). ,
+-+-
,
, ( ) ~ .
, , () . ,
, ,
. , - , . , ,
, 1.
. -
1 « » « » ,
«
»,
u. , , « ».
. 8.5. 7 - S cor ~( » .}.
. ,
,
. , , , .
(. . 10). .
« - »
,
, « - ». , « » (. 8.5),
, ~ (. 8.6), .
.
.
.
. 8.6. () (S. S:;:) 'J « ». , , rb (S.); ra -
S2'
( ).
, .
.
, . 8.7, (, ).
() - S- . ,
, ,
Lys Met. ,
. ,
,
. octatkob-Ls Met (. 8.7).
,
«» (. 8.8). () ( )
. - (. 8.8,) , . (. 8.8,)-
Enz + S Enz S
() (, ).
~ ::e:::--:rtD ~ ~
. 8.9. . (), (). (), (). -
~OM (1) (F.).
. . ,
, «»
,
, . , .
Q ,
. . .
8.9. ,
. ,
. ,
, () .
,
(. 6) (. 9).
, , , ,
, . ~-l ,4- N (. . 13 33),
. ,
: ll 81
Trpl3 Ile 98
Asp 52[ Glu 35 Ala 110
. 8.10. , ,
. " F. , ,
8 II .
.
(. 15000) , 129 . , , ,
. , - .
J. Biol. Chem. 1968: 243, 1663. , , ~ ,
(. 8.10). , ,
D ; Asp 52 Glu 35. , , JO ., Asp 52. st ,
.
. .
,
21
. (. . 5.7.)
, JO ,
KOTOPYJO JO , His 12 His 119. ,
. JO (. 8.11).
(.
8.1). , . .
d , , .
,
JO :
, , , .
82 8
8.1. ~t (S) () . (. 3). ( Dayhoff . .. ed.: Atlas of roti Sequencc and Structure. Vol. 5. National Biomedical Research Foundation. 1972. .)
(S)
. ~
100. Q.o. 100 (QIO = 2) , . 10- . (.
)
QIO' .
. 8.12. .
.
.
, ~eHTa ,
( ).
. .
,
.
, .
.
, .
,
5,0 9,0. . . .
.
1. .
2. . , , ~l . (Enz-) (SH +):
Enz- + SH+ - Enz - SH.
Enz- + + - EnzH.
SH+ -S + +.
SH + Enz-, Enz- SH+ ,
(. 8.13). , . Enz, S, "" .
8lt8
O~------------------~
. 8.12. '\ .
: u 83
.
, ; (. 6). ,
, , , ll ,
- .
,
, .
:
+ -.
,
. , .
. · 1.
.
[][][],
[][]2.
, n ,
+ - n,
lI , ( ,
n )
n," - +
+ ~ n.
: n <t "",.
,
k, .
CTb1) ( 1)
1 = k,[A]"[B]"'. 1 = k ,[nn,].
, , :
1 = _ l' k,[A]"[B]m = k 1 [","].
k, k _1 I CTporo , Keq • ~, -
, . ,
84 8
1. . k,/k_ ,.
2. ( ) .
. .
.
"m• . 4. (' ,
"m• .
(~GO)
fiG
~G~ = - RT ln ~'
R - . - . .
• A-G . , . . . .
( ). .
. . . .
.
, . .
(6G .. -; . ). , 6GO. I- ,
Jl A-G". - , .
l
.
,
. . (. 7).
, ,
. , . , ,
. ,
.
.
. , . .
(1:). II Il IEnzl (. 7).
l\J jJ , , - Enz - S, r . k .
Enz + S ~ Enz + . k_ 1
. [Enz]:
CKOPOCTb1 = k,[Enz][S]. _1 = k _ ,[Enz][P],
[Enz] :
= ~ = [Enz] [] [] eq k- l [Enz] [S] = [S]·
, . , -
(
6GO). . , ()
, K cq 6G
. . ,
. . . . .
. . [S]
v (. , )
'/Jn,. (. 8.14).
uu , l0 .
ullu
. ,
. . .
1000 .
ll. 85
r~1 . IIlii II.
100 I /, 1000 .
:
[Enz] = 0,1 j = 10-9 , [S] = 0,1 / = 10-3 ,
. .
106. [S] 100 ,
10 000 .
. , . 8.14, . 8.15. , , . ,
Enz + S Enz - S ( Enz - S) , . ,
IS) nz, S (. . Enz - S), v IS). , -
[S], Enz S, - ,
.
, d
.
( V maxl2), .
-
, . {" [S] (.
8.14). . .
[S] , v [S]; , .
•
.
-ll
. ,
[S] .
. 8.15. . 8 () () . . (). , , , . ·
. 8.14.
86 8
1. IS) ( . 8.14 8.15). [S] , . , Vmax ,
. , :
m [S] = .
.'1 ,
J1 Jl0 (. . ' ), v [S].
2. ISI ( . 8.14 8.15). , . .
Vmax [S] v= .
, r IS), , v V max'
3. 151 = "'I ( . 8.14 8.15).
Vmax [S] {' = . Vmax [S] Vmax [S] + [S]'
v= = = [S] + [S] 2 [S] 2 .
, ,
, v . :
, .
Vmax (
, ) v [S] . ,
, . .
:
, ,
, -
=+,
= l/v = l/[S). (.. l/v)
(.. 1/[S), , , I/Vmax ' KM/Vmax ' , ( ), , .
=--=--.
-
(. 8.16). , , , , .
[S] . , . v , IEnz.). , , , "
- , ,
. ,
[S]. , [S), .
--.,
. 8.16. - ( Ijvx I/[S),
V '
: 87
V j I V~ax -= -V-+--. [S]
Vmax, v/[S] ( ) V ( ). , , VmaJKM , , ,- Vmax' beh-I/ ·
- - ,
Vmax .
. , 100 , ,
(VmaJ . . , r
Vmllx ' , , .
Kd - -
, Kd Enz--S:
k 1
k_ 1 K d =-,;:.
- , .
Kd ~
. · .
, ,
- . ,
k
. ,
Enz - S Enz + S ,
u + : k
Enz-S d Enz + . k -2
~- . -
[ Vmax S], V = 2'
1
I/ = I/Kd
. k2 + k_ 1 '* k l' l/KM
.
, --
, (. . 6 10), - . fJ [S] (. 8.17). - .
(. 6).
v
O~~ _________________________ ~
88 8
, :. IVI
, (
).
, 02 (. 6). , , ,
t' log = n log [S] -Iog k',
VmaJl - t'
k' - . .
, [S] k', n- [S). . 8.18 , ,
. log (v/Vma .. - v) log [S] , n, n ,
. n = 1 . n > I ; n. .
n < 1 .
(v = V /2), v/( Vma .. -v) = 1 log [v/( Vma .. - - v») = . , Sso ( , ),
, log [t,/( Vmax-V») = , .
v log
Sso
. 8.18. . .1, , -
.
.
- - , (
) ( ) . ,
.
. , ( ), .-
'r (
).
(l) .
, (1), (S). , Enz - S Enz - 1, . . - . ~
, ,
. -
(1). (S).
-
(. 8.19). ( 00C~H2~00 ) , Enz - 1. - .
Enz --:- 1 .
" Enz-I ~ Enz + 1
1 '
~ Enzl (- ~ Enz + Enz .) ~ zS(.)'" Enz +
- -
Enz-S. , 1 (, ). (Enz), S, Enz-S, Enz + , . , ( ) .
(; ) . ,
I S. Enz - S Enz-I. [Enz-S)/[Enz-I] . ~
S Enz - 1 . , 1 (. 8.20).
1
[S) ( Oj[S).
"'1
. 8.20 - , -. (v) S . ,
, .
. , 1/ Vm~x; ,
S (1/IS) = ) v , . ,
( ).
(:-- 1/ ' < -1/ )' , )' (' ) . , ,
1. 3 K j • I ,
[1] .. , ( ) , . , .
. ,
, , .
, S 1 . S , ,
. ,
( Vmax)' , , • 1 S ,
Enz-I, Enz-IS. Enz - IS , ,
Enz-S; , . , -
90 l:.8
Enz +
. 8.2] ]/v ]j[S] ( , I lI).
. .
.
, ,
,
.
. , , -
1 [s]
. .2). )-·) .'1 n HCK~lKYPCH fHnro Rtl.
, .
(
)
. ll .
, (
). «» - ,
.
,
: , , , , , . ,
, «» «» . aKT~HOCT
(. . 6). ,
, ,
. 10 .
Christensen . N. Dissociation, Enzyrne Kinetics, Bioenergetics, Saunders, 1975.
Eng/e . . Enzyme Kinetics, Wiley. 1977. Piszki»'icz D. Kinetics of Chernical and Enzyrne-Catalyzed Re
aetions, Oxford Univ. Press, 1977. Segel 1. . Enzyrne Kinetics. Wiley, 1975. Sigman D .. S., Mooser G. Chernieal studies of ~rn active si
tes, . Rev. Biochern., 1975, 44, 889. Van Ta"'!len .. (ed.) Bioorganie Chernistry, Vol. 1, Enzyrne
AetlOn, 1977; Vol. 2, Maero- and Multirnoleeular Systerns, 1977; Vol. 3, Substrate Behavior. 1978, Academic Press.
9
. , n
, .
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