cysteine palmitylation

Christoph Turck (turck@itsa.ucsf.edu)
Thu, 30 Apr 1998 11:26:05 -0700 (PDT)

We want to map a cysteine palmitylation site in a protein. The protein is
modified to about 50 % and also spiked with tritiated palmitylated
protein.
Among other things we are planning to subject the protein to CNBr cleavage
and 1 % Tfa to elute it from an affinity resin. Does anybody know if the
thioester bond is stable under these conditions?
We are also debating an in-gel tryptic digest followed by RP-HPLC of
the tryptic peptides and mass-spec and Edman degradation (cold and hot). I
am a little worried that the rather hydrophobic palmitylated tryptic
peptide won't elute from the gel. I also don't know how well these
peptides behave in electrospray mass spec.
Any suggestions on the above would be welcome.

Thanks in advance,

Chris (turck@itsa.ucsf.edu)