Re: Protein Sequencing

scott_leigh@cohesiontech.com
Tue, 30 Mar 99 07:43:53

I've got a HP241 sequencer as well. Although I don't see methionines
very often, when I have, they have been very clean, no sign of a peak
coeluting with PTH-lysine, even at about 200 - 300 pmol of
PTH-methionine. Don't know what to tell you, maybe an old reagent
issue.



Scott Leigh, PhD
Cohesion Technologies
2500 Faber Place
Palo Alto, CA 94303
(650) 354-4847
scott_leigh@cohesiontech.com
sleigh@cson.com


______________________________ Reply Separator _________________________________
Subject: Protein Sequencing
Author: trobinson@pdl.com (Tom Robinson) at INTERNET
Date: 3/29/99 4:36 PM

I have a question about N-terminal protein sequencing. The known
N-terminal sequence of the protein I am sequencing is DIQM.... In the
fourth cycle I see the expected PTH-methionine peak, but I also see a
smaller peak eluting at the retention time of PTH-lysine. I get about 500
picomoles of methionine and 100 picomoles of "lysine" in cycle 4. I am
using an HP 241 sequencer. I have tried sequencing another protein with a
methionine residue and observed the same thing. Can methionine be
converted to lysine or something that elutes at the same time as lysine by
the cleavage/conversion chemistry? Does anyone know the cause of this?
Could old reagents cause this?
Thanks,
Tom

Tom Robinson
Protein Design Labs, Inc.