[ccp4bb] Mercaptoethanol

2009-05-12 Thread Wataru Kagawa

Hi.

We are trying to prepare heavy atom derivatives of a protein in which  
a surface residue was mutated to cysteine. Mass spectrometry of the  
purified cysteine mutant showed an additional peak with a molecular  
mass that correspond to a mercaptoethanol-bound cysteine mutant. I am  
wondering whether the bound mercaptoethanol prevents the heavy atom  
from binding to the cysteine residue.


Any information would be greatly appreciated.

Wataru Kagawa


Re: [ccp4bb] Mercaptoethanol

2009-05-12 Thread Wataru Kagawa

Hi Bill:

Thanks for the prompt reply. I will try purifying the protein without  
mercaptoethanol.


Wataru


On 2009/05/13, at 11:42, William G. Scott wrote:


HI Wataru:

I think it makes an R-S-S-R' bond, so the disulfide will prevent the  
heavy atom from binding, unless exchange is rapid.


I hope all is well.

Bill


On May 12, 2009, at 7:17 PM, Wataru Kagawa wrote:


Hi.

We are trying to prepare heavy atom derivatives of a protein in  
which a surface residue was mutated to cysteine. Mass spectrometry  
of the purified cysteine mutant showed an additional peak with a  
molecular mass that correspond to a mercaptoethanol-bound cysteine  
mutant. I am wondering whether the bound mercaptoethanol prevents  
the heavy atom from binding to the cysteine residue.


Any information would be greatly appreciated.

Wataru Kagawa