I got very efficient cleavage at the carboxyl terminus of leucine under
almost the same conditions, except that the ammonium bicarb was 25 mM,and
contained 5% acetonitrile. The only published results I have are shown in a
paper in Cell.
C.J. Aldrich, A. DeCloux, A.S. Woods, R.J. Cotter, M.J. Soloski and J.
Forman. Identification of a Tap-Dependant Leader Peptide Recognized by
alloreactive T Cells Specific for a Class Ib Antigen, Cell 79 (1994)
649-658.
Amina
Amina S. Woods, Ph.D.
NIDA Intramural Program, NIH
5500 Nathan Schock Drive
Baltimore, MD 21224
Tel: 410-550-1507
Fax: 410-550-2971
e-mail: awoods@intra.nida.nih.gov
-----Original Message-----
From: Association of Biomolecular Resource Facilities
[mailto:abrf-request@aecom.yu.edu]On Behalf Of Ioannis A. Papayannopoulos
Sent: Friday, June 23, 2000 7:45 AM
To: Recipients of ABRF List
Subject: Re: Chymotrypsin
In my experience TLCK treated chymotrypsin (from bovine pancreas -- I am
not aware of commercially available enzyme from any other source) will cut
with high efficiency after Leu (but usually not between Leu-Pro) under the
"standard" digestion conditions, e.g. 0.1 M ammonium bicarbonate, 0.1 mM
calcium chloride, pH 8.0, 37oC, for anywhere from 2 hrs to overnight,
depending on your protein. Under these conditions, chymotrypsin will also
cut after Phe, Tyr, and Trp, and, to a lesser extend, after Met and several
other amino acids, non specifically.
Ioannis Papayannopoulos
AstraZeneca R&D Boston
(Still in) Worcester, MA
Jun Ouyang <Jun.Ouyang.B@bayer.com> wrote:
Would some experts share experience with me that under what condition
chymotrypsin will definitely cut after Leucine?
Thanks in advance.
Jun Ouyang
Bayer Corp.
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