Dear Bernhard，
Thank you for your reply. there is No His amino acid in N-ter 30 amino
acids, we have suspected Zinc Finger in this N-Ter,add ZnSO4 to Coordinate
during induced .Also add DTT/β-Me in Buffer During Q/S,SEC etc.All in
all,Appreciate your kindly advices!
Best Wishes
Guohui,SHANG
-----原始邮件-----
发件人:"Bernhard Lechtenberg" <[email protected]>
发送时间:2020-09-15 14:49:39 (星期二)
收件人: [email protected]
抄送:
主题: Re: [ccp4bb] A Problem of my Research Protein
Hi Guohui,
Are you sure this is caused by disulfide bond formation? Do you have any
reducing agent in your buffer? 5 Cys residues in 30 amino acids looks very much
like a zinc finger to me. Is there any evidence for that? Are there any other
zinc binding residues, i.e. His in this region?
Good luck,
Bernhard
From: CCP4 bulletin board <[email protected]> on behalf of Guohui SHANG
<[email protected]>
Reply to: Guohui SHANG <[email protected]>
Date: Tuesday, 15 September 2020 at 4:22 pm
To: "[email protected]" <[email protected]>
Subject: [ccp4bb] A Problem of my Research Protein
Hi Everyone，
Well,My Research Protein is easily Dimerzation caused by Disulfide bond
for many Cys.And it leads to the SEC peak too wide about 5 mL. Then I Cut N-ter
30 Amino Acids in which 5 spaced Cys,but the SEC Peak is still wide and hard to
Crystallize（as the protein is very pure and clean）.Besides, I have tried
different Buffer pH ＜ pI or pH ＞ pI，but the SEC peak Do not Work at all.
Anyone could offer your kindly ideas,I would thank you very much！
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