Tryptic digest of the excised gel band, followed by rigorous peptide matching helped us identify a fragment in a very recent case. The crystallographic results confirmed MS findings.
Curiously, the ends were 'ragged' so direct ESI-MS was not very useful, likewise the N-terminal sequencing would not have been a good idea because of the heterogenous N-terminus. Artem -----Original Message----- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Klaus Futterer Sent: Tuesday, July 01, 2008 7:11 AM To: [email protected] Subject: [ccp4bb] Sequence of crystallised protein fragment We have a 150 kDa protein that reproducibly crystallises at one of the Hampton Screen conditions. However, we know from SDS gel analysis that the crystals contain only a 45 kDa fragment, that forms through proteolytic cleavage over time. We would like to determine the sequence boundaries of the fragment. We believe a combination of N-terminal sequencing plus MS analysis might give us the information we need, but I was wondering whether the ccp4bb community has other suggestions. Klaus --------------------------------------------------------------------- Klaus Fütterer, Ph.D. School of Biosciences P: +44-(0)-121-414 5895 University of Birmingham F: +44-(0)-121-414 5925 Edgbaston E: [EMAIL PROTECTED] Birmingham, B15 2TT, UK W: www.biochemistry.bham.ac.uk/klaus/ ---------------------------------------------------------------------
