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Re: [ccp4bb] putting in methionines for SeMet crystals |
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CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999Subject: Re: putting in methionines for SeMet crystals From: Charlie Bond Charles {- dot -} Bond {- at -} UWA {- dot -} EDU {- dot -} AU Date: 2009-02-03 We had success substituting an Arg to Met. It might seem counterintuitive (basic -> hydrophobic), but the underlying reason was that this Arg was involved in DNA-binding and the mutant was designed to effectively clip off the guanidino group while leaving the rest of the sidechain intact. It appeared to work well and in the process added a Met to our sequence which helped MAD phasing once we got crystals. Cheers, Charlie > amit sharma wrote: >> Dear All, >> >> I have a 9-kDa protein that crystallizes well. Since there is no >> structural homologue for this molecule, I intend to make Se-Met >> derivative of the protein. The molecule has no Met/Cys residues in its >> sequence. I wanted to know where in the sequence should I mutate, so >> that the folding/crystallizability of the protein is not compromised. >> >> Any suggestions would be of great help. >> >> Thanks in advance, >> -- >> Amit Sharma, Ph.D. Research Associate, Department of Biology, >> University of York, YO10 5DD UK > -- Charlie Bond Professorial Fellow University of Western Australia School of Biomedical, Biomolecular and Chemical Sciences M310 35 Stirling Highway Crawley WA 6009 Australia Charles.Bond@uwa.edu.au +61 8 6488 4406 CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999 |
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