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.b...@uwa.edu.au
+61 8 6488 4406
