Hello,
I am writing to see whether anyone has experience in digesting actin (42
kDa of monomeric bovine muscle). It contains 6 Cys residues, some of
which may be modified by oxidized glutathione (GSSG). My goal is to
identify the number and sites of glutathionylation. I have tried
in-solution digestion by trypsin, chymotrypsin, Glu-C, Lys-C and CNBr.
The sequence coverage is quite good. It seems that trypsin and Lys-C
gave the most fragments, however, there is a lot of incomplete digestion
giving rise to large peptides. I often see peptides in the 4000-8000 m/z
range that did not match any predicted masses. I thought that one
possibility is disulfide linkages, however, they did not match any of my
observed masses either. I would like to improve digestion until I get
more complete cleavage. I also looked into literature and there is not a
lot to refer to. I'd appreciate some help if anyone has experience in
actin.
Sincerely,
Emily Boja
email: bojae@nhlbi.nih.gov
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