Clostripain

Lee, Karen (klee@Genzyme.com)
Thu, 15 Oct 1998 14:04:07 -0400

We are currently generating peptide maps of a recombinant protein using
Clostripain from Promega. These maps are very reproducible and cleavage
is always complete. The one problem we are having is very consistent
cleavage of our protein at Arg and Lys rather than at Arg alone. We've
confirmed these results using LC/MS on a number of different occassions
and have altered the digest conditions (enzyme/protein ratio, incubation
time and temp etc.) but still see consistent cleavage at Lys. In fact,
our Clostripain maps are very similar to tryptic maps of the same
protein! I have contacted Promega and they have no information or
history relating to Clostripain and cleavage at Lys residues. Does
anyone have any suggestions or experience with this or should I start
digging out my succinylation protocols?

Thanks for your help!

Karen

Karen L. Lee
Research Scientist
Genzyme Corporation
1 Mountain Rd.
Framingham, MA 01701-9322
Phone: (508) 872-8400
Fax: (508) 872-9080
E-Mail: KLee@Genzyme.com