I don't know that this info will be of any help. A long time ago I
converted phosphoserines to S-ethylcystein, using ethanethiol in 1.0 M
NaOH, then did several enzymatic digests using Endoproteinase-Lys C and
Glu-C as well as trypsin, and had no problems what so ever. I used 25mM
ammonium bicarbonate pH 7.8 for buffer in all cases, and a 1:50
substrate:Enzyme ratio. I don't know if you are converting phosphoserine,
but if you are you should read the following paper:
Holmes, C.F.B. FEBS Lett. 215 (1987) 21-24.
Using ethanethiol even in the hood will make the lab stink for days.
Amina
Amina S. Woods, Ph.D.
Johns Hopkins School of Medicine
725 N Wolfe street, Baltimore, MD 21205
Biophysics Building Rm. B7
Tel: (410) 614-4981, Fax: (410) 955-3420
e-mail: awoods@jhmi.edu
----------
From: Daniel Hess[SMTP:Daniel.Hess@fmi.ch]
Sent: Thursday, June 24, 1999 3:56 AM
To: Recipients of ABRF List
Subject: S-aminoethyl cystein
Dear ABRFrs
does someone have a protocol for the modification of Cys to S-aminoethyl
cystein for cleavage with Lys-C? Supplier of the reagent? Conditions?
Thanks for any help!
Greetings Daniel
Dr. Daniel Hess
Friedrich Miescher Institut
Postfach 2543
CH-4002 Basel
Tel.: 061 697 85 44 or 061 697 68 68 FAX: 061 697 3976
e-mail: dhess@fmi.ch