Dear all,
I have contacted a few of you personally based on the archives of this
server, and I thank you for your efforts, but I still can't solve my
problem of C-terminal sequencing. Perhaps the group at large will be able
to offer their advice.
I have a bacterial protein of around 100kDa that I believe is processed
(several amino acids are cleaved off by an active serine protease
catalytic site 800 residues upstream) at the C-terminus. I want to be
able to determine the site of processing. So I am on the search for a
facility who can perform this analysis.
The protein can be electroblotted to either PVDF or nitrocellulose or cut
out of a gel. Sorry, but it's not purified to 100% (it is around 80%
pure). Someone mentioned using mass spect to do the analysis, but it may
not be reliable for such large proteins. Tryptic digests also may not
gaurantee results if the C-terminal fragment does not elecrtospray.
C-terminal sequencing would be ideal.
Many of the resources mentioned in previous e-mails no longer offer the
service or are unwilling to take on new outside projects. The
facility at the Univ. of Michigan was mentioned several times, but neither
the e-mail addresses nor the phone lines working at this facility.
I would greatly appreciate any input from anyone who can provide this
service or knows of someone who still does. Please reply to me personally
at pdutta@umaryland.edu . I don't know if I'm correctly registered on
the list serv yet.
Thank you.
--Pinaki Dutta
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