Dear James:

The problem is that the equation is for an equilibrium constant, and  Km is not 
one.  Kd is one.  If you can measure Kd, what you describe I think is fine.  If 
you are measuring Km, you have to find a way of justifying the Km ~ Kd 
approximation, which usually is only safe for enzymes that turn over very 
slowly.

I've never done these kinds of experiments, so someone else needs to answer 
this.  Sorry.  I've always been content to push the deltaG button in COOT.

All the best,

Bill


On Nov 18, 2009, at 10:18 AM, james09 pruza wrote:

> Dear Sir,
> 
> At equilibrium, how can one determine delH and delS, if temp. and km are
> known.
> 
> I have km value and temperatures. I have to calculate delH and delS in
> equilibrium.
> Should I plot *lnKm vs 1/T* to have a slope of -delH/R and intercept of
> delS/R. Multyplying this value with R, will give the value of delH and delS
> , respectively.
> If delG= delH - TdelS
> delG= -RTlnK
> 
> So, -RTlnK = delH -TdelS
> 
> lnK = -delH/RT + delS/R
> 
> Please suggest If I am wrong. I have planned to do ITC with the mutants as
> well to get the perfect parameters.
> Thanks for help.
> 
> On Wed, Nov 18, 2009 at 7:44 PM, William G. Scott <
> wgsc...@chemistry.ucsc.edu> wrote:
> 
>> The K is an equilibrium constant.  Km is not an equilibrium constant.  It
>> is a collection of rate constants. Kd is an equilibrium constant, but you
>> have to be careful to make sure that the enthalpy and entropy terms
>> correspond directly to this as well. The equation you quote implies standard
>> state.
>> 
>> Km is not the reciprocal of Kd.  It is not an equilibrium constant.
>> 
>> 
>> On Wed, Nov 18, 2009 at 8:24 AM, james09 pruza <james09x...@gmail.com>wrote:
>> 
>>> Dear All,
>>> Sorry for the non-ccp4 query.
>>> I have solved a crystal structure of an enzyme and woring on its
>>> biochemical aspect. We have a mutant of this enzyme and we are comparing
>>> some thermodynamic parameters of this enzyme with mutant( lke delH and delS,
>>> delG). we have done the expt at different temp. and know the km value at
>>> these temp. Now the question is how to get the value of delH and delS.
>>> The relation of these parameters is:-
>>> -RTlnK = delH - TdelS
>>> 
>>> Is the K in this relation is km or kd. If it can be treated as kd, so it
>>> should be 1/km.
>>> 
>>> All suggestions are welcome.
>>> Thanks
>>> James
>>> 
>> 
>> 
>> 
>> 

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