lecture of graduiertenkolleg - helms

50
1. Lecture SS 2006 GK 1276 1 Lecture of Graduiertenkolleg - Helms V1: Reaction rate theory V2: Kramer‘s theory V3: Reaction rates for electronic transitions V4: Potential and free energy landscapes V5: Lattice optimization V6: Optimization methods in protein folding V7: Protein folding with molecular dynamics V8: Manipulating potential energy landscapes with forces

Upload: thina

Post on 06-Jan-2016

31 views

Category:

Documents


0 download

DESCRIPTION

Lecture of Graduiertenkolleg - Helms. V1:Reaction rate theory V2:Kramer‘s theory V3:Reaction rates for electronic transitions V4:Potential and free energy landscapes V5:Lattice optimization V6:Optimization methods in protein folding V7:Protein folding with molecular dynamics - PowerPoint PPT Presentation

TRANSCRIPT

Page 1: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 1

Lecture of Graduiertenkolleg - Helms

V1: Reaction rate theory

V2: Kramer‘s theory

V3: Reaction rates for electronic transitions

V4: Potential and free energy landscapes

V5: Lattice optimization

V6: Optimization methods in protein folding

V7: Protein folding with molecular dynamics

V8: Manipulating potential energy landscapes with forces

Page 2: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 2

V1: Chemical Kinetics & Transition States

see chapter 19 in book of K. Dill

Aim: describe kinetics of processes on energy landscapes

(e.g. chemical reactions).

- detailed balance- mass action law- temperature effect, Arrhenius law- concept of transition state/activation barrier transition state theory- -value analysis- effect of catalysts

Page 3: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 3

Rate theory

Rate theory provides the relevant information on the long-time behavior of

systems with different metastable states important for understanding of many

different physical, chemical, biological, and technical processes.

Arrhenius (1889)

Wigner (1932), Eyring (1935) Transition State Theory (TST)

Pechukas (1976): proper definition of the transition state

Chandler (1978): the activation energy is a free energy

Kramers (1940): effect of friction on reaction rates

Pollak (1986): link of Kramers‘ expression to TST

Pollak, Talkner, Chaos 15, 026116 (2005)

Page 4: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 4

Reaction rates are proportional to concentrations

B

k

k

A

r

f

Lets consider a simple kinetic process, the interconversion between 2 states,

kf and kr : forward and reverse rate coefficients.

How do the amounts of A and B change with time t,

given the initial amounts at time t = 0 ?

tBktAkdt

tBd

tBktAkdt

tAd

rf

rf

The two equations are coupled.

One can solve them by matrix algebra …

Page 5: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 5

Excursion: coupled differential equations

yxy

yxx

2

4

y

x

y

x

12

14

Matrix diagonalisation can be used to solve coupled ordinary differential equations.

For example, let x(t) and y(t) be differentiable functions and x' and y' their time derivatives.

The differential equations are relatively difficult to solve:

By diagonalizing the square matrix, we get

but u' = ku for k = const is easy to solve.

The solution is u = Aekx where A = const.

translate the ODEs into matrix form

y

x

y

x1

21

11

20

03

21

11

www.algebra.com

Page 6: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 6

Excursion: coupled differential equations

y

x

y

x1

21

11

20

03

21

11

y

x

v

u1

21

11

y

x

v

u1

21

11

By diagonalizing the square matrix, we get

We then put It follows that

Thus

v

u

v

u

20

03

The solutions of this system are found easily: t

t

Dev

Ceu2

3

with some constants C and D.

y

x

v

u1

21

11

y

x

v

u

21

11With

tt

tt

DeCey

DeCex23

23

2

Page 7: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 7

Reaction rates are proportional to concentrations

tAkdt

tAdf

tkA

tAdtk

A

dAf

t

f 0ln

0

tk feAtA 0

With this technique, we could solve our system of coupled diff. equations.

In the special case that kr << kf, the first equation simplies to

If [A(t)] + [B(t)] = constant, then

tk feAtB 0constant

Page 8: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 8

[A]eq and [B]eq : equilibrium concentrations.

To see that this is a condition of equilibrium follows from inserting

into

resulting in

At equilibrium, rates obey detailed balance

eqreqf BkAk

eqreqf BkAk

tBktAkdt

tBd

tBktAkdt

tAd

rf

rf

The principle of detailed balance says that the forward and reverse transitions must

be identical for an elementary reaction at equilibrium:

Taken from Dill book

0,0

dt

tBd

dt

tAd

Page 9: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 9

At equilibrium, rates obey detailed balance

r

f

eq

eq

k

k

A

BK

The detailed balance condition relates the rate coefficients kf and kr to the

equilibrium constant K:

Taken from Dill book

For more complex systems, the principle of detailed balance gives more information beyond

the statement of equilibrium. For a system having more than one elementary reaction, the

forward and reverse rates must be equal for every elementary reaction.

For this system:

Let‘s consider a 3-state mechanism with

kIA 0, kBI 0, kAB 0.

AB

BA

eq

eq

BI

IB

eq

eq

IA

AI

eq

eq

k

k

B

A

k

k

I

B

k

k

A

I ,,

Page 10: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 10

These are two independent equations for 3

unknown concentrations

the system has an infinite number of

solutions.

In mechanism (b), all rates of the

Denominator in

are zero mechanism (b) is impossible.

At equilibrium, rates obey detailed balance

BAIB

IBAI

kBkI

kIkA

This results in the mechanism shown right.

The only conditions for equilibrium are:

BIABIA

IBBAAI

IBBAAI

BIABIA

BI

IB

eqAI

IA

BA

ABeq

BA

ABeqeqeq

IA

AIeq

AB

BA

eq

eq

BI

IB

eq

eq

IA

AI

eq

eq

kkk

kkk

kkk

kkk

k

k

Ak

k

k

kA

k

kABA

k

kI

k

k

B

A

k

k

I

B

k

k

A

I

1or,1

or)2(

)3(,)1(

,,

Taken from Dill book

Page 11: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 11

At equilibrium, rates obey detailed balance

The principle of detailed balance says that forward and backward reactions at

equilibrium cannot have different intermediate states.

That is, if the forward reaction is A I B,

the backward reaction cannot be B A.

Page 12: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 12

The mass action laws describe mechanisms in chemical kinetics

Suppose the following reaction leading from reactants A, B, and C to product P:

PcCbBaA In general, the initial reaction rate depends on

- the concentrations of the reactants

- the temperature and pressure

- and on the coefficients a, b, and c.

Kinetic law of mass action (CM Guldberg & P Waage, 1864):

„the reactants should depend on the stoichiometry in the

same way that equilibrium constants do“.

cbaf CBAk

dt

Pd

Although mass action is in agreement with many experiments,

there are exceptions.

These require a quantum mechanical description.

Page 13: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 13

Energy Barriers

Where do energy barriers come from?

Why do reactions have activation barriers?

Different processes are characterized by similar energy barriers that are due to

very different mechanisms.

Effects on energy barriers- Chemical reactions - temperature- protein:ligand association - pH

- protein:protein association - D2O vs. H2O

- protein:membrane association - viscosity- protein:DNA association- during protein folding- time scales of protein dynamics- vesicle budding- virus assembly

Page 14: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 14

History of Energy Barriers of Chemical Reactions

1834 Faraday: chemical reactions are not instantaneous because there is an

electrical barrier to reaction

1889 Arrhenius: reactions follow Arrhenius law

with an activation barrier Ea.

Bodenstein: reactions occur via a series of elementary steps where bonds break

and form. Bodenstein showed that Arrhenius‘ law is applicable only to

elementary reactions. Overall reactions often show deviations.

1935 Polanyi & Evans: bonds need to stretch during elementary reactions.

The stretching causes a barrier. Bonds also break.

Physical causes of barriers to chemical reactions

bond stretching and distortion

orbital distortion due to Pauli repulsions (not more than 2 electrons may occupy one orbital)

quantum effects

special reactivity of excited states

kT

Ea

erateRate

0

Page 15: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 15

Energy Barriers of Chemical Reactions

When chemical bonds need to be „broken“,

- nuclei need to move only over small distances,

- electrons need to redistribute „settle down“ in different orbitals intermediate state has high energy

One can compute the energy barriers by electronic structure methods.

However, these calculations do not explain why the barriers arises.

Chemists like to think in concepts and rules and like to separate these.

How fast can a reaction proceed? Even within one bond vibration.

Such processes need to be activated.

E.g. bond length should stretch far beyond equilibrium distance.

This is possible by statistical fluctuations and by coupling with other modes

(large energy becomes concentrated in this mode).

Page 16: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 16

Energy Barriers of Protein:Ligand InteractionStep 1: Protein and Ligand are independently solvated (left picture below)

Step 2: The Ligand may preorganize into its binding conformation (costs usually 3 kcal/mol)

Step 3: The Ligand approaches the binding pocket of the protein.

System partly looses 6 degrees of freedom (CMS of ligand: 3 translation, 3 rotation)

Step 4: The Ligand enters the binding pocket of the protein Waters are displaced from binding pocket.

Sometimes: simultaneous conformational changes of protein/receptor

Receptor

Ligand

Bound and associated H2O

Displaced H2O

No collective modes!

Page 17: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 17

Steps involved in protein-protein association:

- random diffusion (1) + hydrodynamic interaction

- electrostatic steering (2)

- formation of encounter complex (3)

(Possible: large-scale conformational changes of

one or two proteins)

- dissociation or formation of final complex via TS (4)

Origin of Barrier (4): System partly looses 6 degrees of freedom (CMS: 3 translation, 3 rotation) Desolvation: large surface patches need to be partially cleared from water Induced fit of side chains at interface potential entropy loss

Energy Barriers of Protein:Protein Interaction

Effects of hydrodynamicInteractions:(left) effect of translation(right) effect of rotation

Page 18: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 18

Energy Barriers of Protein:Membrane Interaction

Membrane surface either carries a net negative charge (mixture of neutral and

anionic lipids) or has a partially negative character.

Cloud of positive counter ions accumulates near membrane to compensate

membrane charge.

Membrane surface is not well defined and quite dynamic, ondulations.

wikipedia.org

Page 19: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 19

Energy Barriers of Protein:Membrane Interaction

Neutron scattering: four layers of ordered water molecules

above membrane – also found by MD simulation.

Water layers significantly weaken membrane potential.

Lin, Baker, McCammon, Biophys J, 83, 1374 (2002)

Interaction potential of protein:membranesystems is largely unknown.

Page 20: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 20

Energy Barriers of Protein:DNA Interaction

DNA backbone carries strong permanent negative charge.

is surrounded by cloud of positive ions and coordinating water molecules

Protein must displace this cloud and must form very polar interactions with DNA

backbone.

Spatial distribution functions of water, polyamine atoms and Na+ ions around a CA/GT fragment. View of the minor groove. Data are for systems with 30 diaminopropane2+ (A), 30 putrescine2+ (B), 20 spermidine3+ (C) and 60 Na+ (D), averaging the MD trajectories over 6 ns, with three decamers with three repeated CA/GT fragments in each decamer. Water (oxygen, red; hydrogen, gray) is shown for a particle density >40 p/nm3 (except for the Na/15 system, where this value is 50 p/nm3); Spherical distribution function of the polyamine N+ atoms (blue) and Na+ (yellow) ions are drawn for a density >10 p/nm3; polyamine carbon and hydrogen atoms not shown.

Korolev et al. Nucl Acid Res 31, 5971 (2003)

Page 21: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 21

Energy Barriers of Protein:DNA Interaction

Recognition shows Faster-than-diffusion paradox (similar to Anfinsen paradox

for protein folding).

Maximal rate achieveable by 3D diffusion: 108 M-1s-1

This would correspond to target location in vivo on a timescale of only a few

seconds, when each cell contains several tens of TFs.

However:

Experimentally found (LacI repressor and its operator on DNA): 1010 M-1s-1.

Suggests that dimensionality of the problem changes during the search

process. While searching for its target site, the protein periodically scans the

DNA by sliding along it. This is best done if the TF is only partially folded and

only adopts its folded state when it recognizes its binding site.

Slutsky, Mirny, Biophys J 87, 4021 (2004)

Page 22: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 22

Time scales of protein dynamics

Motion Spatial extent (nm)Log10 of characteristic

time (s)

Relative vibration of bonded atoms 0.02 to 0.05 -14 to –13

Elastic vibration of globular region 1 to 2 -12 to –11

Rotation of side chains at surface 0.5 to 1 -11 to –10

Torsional libration of buried groups 0.5 to 1 -14 to –13

Relative motion of different globular regions (hinge bending)

1 to 2 -11 to –7

Rotation of medium-sized side chains in interior

0.5 -4 to 0

Allosteric transitions 0.5 to 4 -5 to 0

Local denaturation 0.5 to 1 -5 to 1

Protein folding 3 to 5 -5 to 2

Adapted from http://www.dbbm.fiocruz.br/class/Lecture/d22/kolaskar/ask-11june-4.ppt

Page 23: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 23

Energy Barriers during protein folding

Peptide chain must organize into particular 3D fold: large entropy loss.

Formation of secondary structure elements formation of hydrogen bonds.

This is almost cancelled by loss of hydrogen bonds with solvent molecules.

Burial of hydrophobic surface free energy gain due to hydrophobic effect.

Charged active site residues must be buried in hydrophobic protein interior

often electrostatically unfavorable

Page 24: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 24

Vesicle budding (right, above) does not occur

spontaneously: would be too dangerous for cell.

Distorting plane membrane costs deformation energy

binding of coat proteins reduces energy cost and gives

natural membrane curvature (right, below).

SNARE proteins help to overcome energy cost for fusion of

membranes.

Energy Barriers during vesicle budding

Page 25: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 25

Energy Barriers during formation of virus capsid

Many individual particles combine into one larger particle

big loss of translational and rotational degrees of freedom

Much hydrophobic surface gets buried between assembling proteins free energy gain according to hydrophobic effect (primarily solvent entropy)

Electrostatic attraction: probably not very significant for binding affinity but

important for specificity.

Page 26: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 26

What is the effect of pH on energy barriers?

At different pH, titratable groups will adopt different protonation states.

e.g. at low pH, Asp and Glu residues will become protonated

salt-bridges (Asp – Lys pairs) in which residues were involved will break up.

proteins unfold at low and high pH.

Page 27: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 27

What is the effect of D2O on energy barriers?

It is a common strategy to compare the speed of chemical reactions in H2O and

in D2O.

The electronic energy profile of the barrier is the same.

But the deuteriums of D2O have a higher mass than the hydrogens of H2O.

Their zero-point energies are lower

they need to overcome a higher effective energy barrier

all chemical reactions involving proton transfer will be slowed down,

typically by a factor of 1.4

This is called the „kinetic isotope effect“ (KIE).

Page 28: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 28

What is the effect of viscosity on energy barriers?

Adding co-solvents in the solvent to increase the viscosity should, in principle,

slow down conformational transitions.

It is often problematic that the co-solvent will also change the equilibrium, e.g.

between folded and unfolded states of a protein.

Page 29: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 29

What is the effect of temperature on energy barriers?

In general, higher temperature will enormously speed up activated processes.

However, we often need to consider free energy barriers instead of energy

barriers. The free energy barriers often change considerably with temperature.

E.g. in a MD simulation of a protein at 500K, the protein residues will more

easily overcome individual torsional energy barriers.

But, after a certain time, the whole protein will unfold.

Page 30: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 30

Reaction rates depend on temperatureConsider a binary reaction in the gas phase:

PBAk2

Suppose that

By definition, the rate coefficient k2 is independent of [A] and [B].

But k2 can depend strongly on temperature.

BAkdt

Pd2

The observed dependence of the reaction rate on the temperature is much

greater than one would expect from just the enhanced thermal motions of the

molecules.

Page 31: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 31

1889 Arrhenius: found temperature dependence of

the rates of inversion of sugar in the presence of acids.

Arrhenius cites van‘t Hoff (1884) for suggesting

e–A/T dependence.

Ea: activation energy

Arrhenius postulated that this relationship indicates the existence of an

„activated sugar“ whose concentration is proportional to the total concentration

of sugar, but is exponentially temperature dependent.

Arrhenius is the father of rate theory

Arrhenius and activated molecules

kT

Ea

erateRate

0

Svante Arrhenius 1859 – 1927Noble price 1903

Page 32: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 32

Arrhenius equation

1889, S. Arrhenius started from the

van‘t Hoff equation for the strong

dependence of the equilibrium constant

K on temperature:

2

ln

kT

h

dT

Kd

and proposed that kf and kr also have van‘t Hoff form

22

lnand

ln

kT

E

dT

kd

kT

E

dT

kdaraf

where Ea and E‘a have units of energy that are chosen to fit exp. data.

Ea and E‘a are called activation energies.

Page 33: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 33

Activation energy diagram

According to Arrhenius, it is not the average energy of the reactants that

determines the reaction rates but only the high energies of the ‚activated‘

molecules.

Taken from Dill book

There are two plateaus, one for the reactants and one for the products.

In between lies an energy maximum (also: transition state or activation barrier)

which is the energy that activated molecules must have to overcome on their

way from reactants to products.

Measuring kf as a function of temperature, and using eq. (1) gives Ea.

Measuring the reverse rate gives E‘a.

Measuring the equilibrium constant versus temperature gives h°.

Page 34: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 34

Population at different temperatures

From

Taken from Dill book

r

f

k

kK it follows

aa EEh '

The figure shows how activation is interpreted according to the Boltzmann

distribution law: a small increase in temperature can lead to a relatively

large increase in the population of high-energy molecules.

Page 35: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 35

Arrhenius plots

Integrating

over temperature T

Taken from Dill book2

ln

kT

E

dT

kdaf

gives:

BkTE

f

BkTE

f

BkTE

f

af

eAAek

eek

ek

BkT

Ek

a

a

a

with

ln

H2 + I2 2HI (open circles)

2HI H2 + I2 (full circles)

Diffusion of carbon in iron

The figures show examples of

chemical systems showing

Arrhenius behavior.

Page 36: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 36

Activated processes

Arrhenius kinetics applies to many physical and chemical processes.

When should one treat a process as activated?

If a small increase in temperature gives a large increase in rate,

a good first step is to try the Arrhenius model.

E.g. breaking of bonds.

Counter example: highly reactive radicals. CNHHHCNH 223

These can be much faster than typical activated processes and

they slow down with increasing temperature.

We now describe a more microscopic approach to reaction rates,

called transition state theory.

Page 37: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 37

The energy landscape of a reaction

An energy landscape defines how the

energy of a reacting system depends

on its degrees of freedom.

E.g. A + BC AB + C

Each reaction trajectory would involve

some excursions up the walls of the

valleys.

When averaged over multiple trajectories,

the reaction process can be described as

following the lowest energy route, along

the entrance valley over the saddle point

and out of the exit valley, because the

Boltzmann populations are highest along

that average route. Taken from Dill book

Energy surface for

D + H2 HD + H

The transition (saddle) point isdenoted by the symbol ‡.It is unstable: a ball placed on thesaddle point will roll downhillalong the reaction coordinate.

Page 38: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 38

Eyring theoryThe Eyring-theory or transition state theory (Theorie des Übergangszustandes) is a

molecular reaction theory. It uses molecular descriptors like the partition function and

describes the absolute rate of chemical reactions.

The reactants are separated from the products by

an activation barrier. The reaction from the reactants

via the transition state to the products proceeds

along a trajectory = the reaction coordinate.

Transition state: point of highest potential energy along this reaction coordinate.

Activated complex: atomic arrangement in the transition state.

The main assumptions of TST are:

- the activated complex exists in an equilibrium with the reactants

- All molecules that reach the transition state from the reactant states leave it in direction of

the products. Recrossings are not allowed.

[taken from Dill book]

Page 39: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 39

where is a prefactor with the dimensions of s-1 for

unimolecular reactions and s-1cm-3 for bimolecular reactions.

Wigner and Eyring: The transition state method

kTEa

ek

In the 1930s it was well established that reaction rates k should be written in the form:

Henry Eyring (1901-1981)

[taken from Dill book]

In „The activated complex in chemical reactions“ (1935),

Eyring gave a heuristic derivation of an expression for

the prefactor based on the assumption of an equilibrium

between the activated complex and reactants.

To obtain the time constant, he postulated, that at the

saddle point, any quantum state perpendicular to the

reaction coordinate reacts with the same universal time

constant kT/2ħ.

The rate is then given by the product of this universal time

constant with the ratio of the partition function of the

activated complex to the partition function of the reactants.

Page 40: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 40

Wigner and Eyring: The transition state method

1932, Wigner also derived an estimate for the tunneling

contribution to the thermal flux of particles crossing a

barrier.

1932 Pelzer & Wigner: estimated rate of conversion of parahydrogen into normal

hydrogen.

To compute the reaction rate, they use a thermal equilibrium distribution in the vicinity of

the saddle point of the PES and estimate the unidirectional classical flux in the direction

from reactants to products

They ignore the possibility of recrossings of the saddle

point noting that their probability at room temperature

would be rather small.

How can one define the „activated complex“?

Eyring‘s definition is questionable.

Wigner‘s definition leaves no ambiguity: the best dividing

surface is that which minimizes the unidirectional flux from

reactants to products. Eugene Wigner (1902-1995)Noble price 1963

Page 41: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 41

The transition state

(left) Contour plot of a reaction pathway (- - -) on an energy landscape

for the reaction A + BC AB + C. The broken line shows the lowest-energy

path between reactants and products

(right) The transition state is an unstable point along the reaction pathway

(indicated by the arrow) and a stable point in all other directions that are

normal to the reaction coordinate.

Taken from Dill book

Page 42: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 42

Calculating rate coefficients from TST

Let us consider the reaction

by transition state theory:

Divide the reaction process into two stages:

(1) the equilibrium between the reactants ant the transition state (AB)‡ with

‚equilibrium constant‘ K‡

(2) a direct step downhill from the TS to the product with rate coefficient k‡:

PBAk2

PABBA kK ‡‡ ‡

Key assumption of TST: step (1) can be expressed as an equilibrium between

the reactants A and B and the transition state (AB)‡ , with

BAB

KA

‡‡

even though (AB)‡ is not a true equilibrium state.

Page 43: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 43

Calculating rate coefficients from TST

The overall rate is expressed as the number of molecules in the TS, [(AB)‡],

multiplied by the rate coefficient k‡ for the second product-forming step

BAKkABkdt

Pd ‡‡‡‡

Because the quantitiy K‡ is regarded as an equilibrium constant, it can be

expressed in terms of the molar partition functions:

kTD

BA

AB eqq

qK

‡‡

where D‡ is the dissociation energy of the TS minus the dissociation energy of

the reactants.

q(AB)‡ is the partition function of the transition state.

Page 44: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 44

relation between value analysis and TST

Later we will characterize the effect of a protein mutant by its -value

0

ln

G

kkTk

wt

mutB

G0 reflects whether the mutant stabilizes the folded state F over the unfolded

state U stronger or weaker than wild-type protein.

According to TST, both wild-type and mutant folding proceed via transition

states with activation free energies G‡wt

and G‡mut.

wtFU

mutFU GGG 0

‡‡‡;‡

‡‡

wtmutkTG

kTGG

kTG

kTG

wt

mut

kTGmut

kTGwt

GGGe

ee

e

k

k

ek

ek

wtmut

wt

mut

mut

wt

A -value of 1 meansthat G0 = G‡ forthis mutant the mutant has the sameeffect on the TS structureas on the folded state this part of the TS structure is folded asin the folded state F.

Page 45: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 45

Catalysts speed up chemical reactions

Taken from Dill book

Free energy barrier G‡ is reduced by a catalyst C.

Catalysts affect the rates of chemical reactions; e.g. enzymes accelerate

biochemical reactions.

Enzymes can achieve remarkable accelerations, e.g. by a factor of

2 x 1023 for orotine 5‘-phosphate decarboxylase.

Linus Pauling proposed in 1946 that catalysts work by stabilizing the

transition state.

PBA k 0

ABC

kc

Linus Pauling 1935

Page 46: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 46

Catalysts speed up chemical reactions

From transition theory we obtain for the catalyzed reaction rate kc (normalized to

the uncatalyzed reaction rate k0)

CAB

ABC

k

kc

0

This ratio represents the ‚binding constant‘ of the catalyst to the transition state

the rate enhancement by the catalyst is proportional to the binding affinity of

the catalyst for the transition state.

This has two important implications:

(1) to accelerate a reaction, Pauling‘s principle says to design a catalyst that

binds tightly to the transition state (and not the reactants or product, e.g.).

(2) a catalyst that reduces the transition state free energy for the forward reaction

is also a catalyst for the backward reaction.

Page 47: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 47

Speeding up reactions by intramolecular localization or solvent preorganization

Taken from Dill book

Reactants polarize, so water reorganizes.

Two neutral reactants become charged in the transition state.

Creating this charge separation costs free energy because it orients the solvent

dipoles.

Page 48: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 48

Speeding up reactions by intramolecular localization or solvent preorganization

Enzymes can reduce the activation barrier by having a site with pre-organized

dipoles.

Taken from Dill book

Page 49: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 49

Funnel landscape describe diffusion and polymer folding

All the processes described sofar involve well-defined reactants and products, and

a well-defined reaction coordinate.

But diffusional processes and polymer conformational changes often cannot be

described in this way. The starting point of protein folding is not a single point on

an energy landscape but a broad distribution.

A bumpy energy landscape, such as occursin diffusion processes, polymer conformationalchanges, and biomolecule folding.

A single minimum in the center may representthe ‚product‘, but there can be many different‚reactants‘, such as the many openconfigurations of a denatured protein.

http://www.dillgroup.ucsf.edu/

Page 50: Lecture of Graduiertenkolleg - Helms

1. Lecture SS 2006

GK 1276 50

Summary

Chemical reactions and diffusion processes usually speed up with temperature.

This can be explained in terms of a transition state or activation barrier and an

equilibrium between reactants and a transient, unstable transition state.

For chemical reactions, the transition state involves an unstable weak vibration

along the reaction coordinate, and an equilibrium between all other degrees of

freedom.

Catalysts act by binding to the transition state structure.

They can speed up reactions by forcing the reactants into transition-state-like

configurations.