trx force tactical kit
DESCRIPTION
TRX ForceTRANSCRIPT
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TRX Force Tactical Kit
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Thioredoxin
protein essential for life in mammals acts as an antioxidant to prevent
oxidative stress acts as an electron donor in a variety
of cellular reactions restores oxidatively inactivated
enzymes to their reduced & active state
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Thioredoxin exists in either the reduced or oxidized form
Trx
HS
HS
Reduced Thioredoxin Oxidized Thioredoxin
LEGEND
SH = thiolS-S = disulfide
Trx
S S
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Thioredoxin participates in dithiol-disulfide exchange reactions
HS S S
S SHS
HS
RNRoxidized
RNRreduced
reduced Trx
oxidized Trx
HS
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Trx
thioredoxin
SH
SH
Trx
S S
ribonucleotidereductase
SH
SH
RNR
S S
thioredoxinreductase
RNRNADPH
NADP+
Thioredoxin reductase & thioredoxinwww.prosuspensiontrainer.com
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Hypothesis Thioredoxin reductase is a major determinant
of thioredoxin redox state If true, then deletion of thioredoxin reductase
should shift the redox state of thioredoxin from primarily reduced to primarily oxidized
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SH
IAA DTT IAM
Protein electrophoretic mobility shift assay (PEMSA)
Iodoacetic acid Dithiothreitol Iodoacetamide
LEGEND
SH = thiol
= Thioredoxin
A- = negative charge
M = neutral state
= DisulfideS
S
acetone wash
SHSHSHSS
SHSHSHSHSHSH
SA-SA-SA-SA-SS
SA-SA-SA-SA-SA-SA-
SA-SA-SA-SA-SA-
SA-SA-SA-SA-SA-
SA- SA-
SA-SA-SA-SHSH
SA-SA-SA-SA-SMSM
SA-
TRXox
TRXox
TRXox
TRXox
TRXred
TRXred
TRXred
TRXred
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Trxred
SHSHSHSHSHSH
Trxox
SHSHSHSHSS
SA-
SA-
SA-
SA-
SA-
SA-
SHSHSHSHSHSH
SMSMSMSMSMSM
SM or SA-
SM or SA-
SM or SA-
SM or SA-
SM or SA-
SM or SA-
DTT
IAA IAMIAA/IAM
Protein electrophoretic mobility shift assay
0
-1
-2
-3
-4
-5
-6
1 2 3 4
TrxSS
TrxSH
urea
PA
GE
-
+
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Representative resultsIA
A onl
y
1 2 3 4 5 37 200
201
229
230
Ladd
er
Wild Type Mutant
Trxoxidized6 7 8 7 9 1
216 2
123
21
Mixed disulfide
12
10
14
13
15
17
17 20
21
18
Trxreduced27
22
27
23
29
29
30 34
36
35
Mut
26%
27%
47%
mixed disulfide
Trxred
Trxox
Trxox
mixed disulfide
Trxred
IAM
onl
y
WT
16%
28%
56%
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Conclusion
Thioredoxin reductase is not the major determinant of thioredoxin redox state
Another pathway must be able to reduce thioredoxin
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Acknowledgement
Dr. Gary Merrill Biochemistry & Biophysics Lab
Dr. Kevin Ahern Howard Hughes Medical Institute Undergraduate, Research, Innovation,
Scholarship & Creativity www.prosuspensiontrainer.com