Hi Jon
there is no surface exposed cysteine nearby which can potentially
make disulfide bond with the modified cysteine intramolecularily.
Dhiraj
________________________________
From: Jon Cooper <jon.b.coo...@protonmail.com>
Sent: Sunday, November 1, 2020 7:41 AM
To: Srivastava, Dhiraj <dhiraj-srivast...@uiowa.edu>; CCP4BB@JISCMAIL.AC.UK
<CCP4BB@JISCMAIL.AC.UK>
Subject: Re: [ccp4bb] Cysteine oxidation product
Hello,
does the protein you added cysteine to (I assume you mean by mutagenesis) have
another cysteine which could make an intra molecular disulphide with it? That
would probably not show on non-reducing SDS?
Best wishes, Jon Cooper
-------- Original Message --------
On 31 Oct 2020, 15:58, Srivastava, Dhiraj < dhiraj-srivast...@uiowa.edu> wrote:
Hi All
I added an extra cysteine residue to one of my protein interactions
partners and somehow it increased the affinity in the absence of reducing agent
however in the presence of reducing agent, the affinity is same. there is no
disulfide bond formation between the two interaction partner as in non-reducing
SDS-PAGE, I see the molecular weights corresponding to only the individual
proteins and not the sum of two.
Does anyone has ever seen this? at pH 8.0, what is the probability that a
surface exposed cysteine will have a net negative charge and still not very
reactive? can it exist as thiolate, sulfinate or sulfonate ion? is there any
such example in literature that anyone knows?
Thank you
Dhiraj
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