In our protein crystallography course, we succesfully used S-SAD phasing
for proteinase K crystals diffracting to 1.58 Å (which is 'low' for these
crystals) at our home source.
I would highly recommend using proteinase K for heavy atom derivative
demonstration purposes as it seems much easier to produce these for
proteinase K than for lysozyme. In our hands quick soaks (i.e. a few
minutes) in 50 mM stocks of e.g. EuCl3 or SmCl3 (Eu works great because of
the strong anomalous signal at the Cu Ka edge) are fine.
Our course is based on course material from UCLA:
http://www.doe-mbi.ucla.edu/~sawaya/m230d/m230d.html
If you are interested, you are welcome to contact me ([log in to unmask]) for
our teaching material.
Cheers,
Thomas
--
Thomas Boesen
Associate Professor, PhD
Centre for Structural Biology
Department of Molecular Biology
University of Aarhus
Gustav Wieds Vej 10C
8000 Århus C
Phone: +45 8942 5257 (lab: +45 8942 5030)
E-mail: [log in to unmask]
web: http://www.bioxray.au.dk/index_uk.php
what is it that you want to demonstrate? If it is just for structure
solution, would S-SAD phasing be an option? That way all you need is a
crystal, probably diffracting to better than, say, 2A. And a little bit of
multiplicity.
I assume this is feasible for Lysozyme whereas I don't know the
diffraction qualities of proteinase K crystals.
Tim
--
Tim Gruene
Institut fuer anorganische Chemie
Tammannstr. 4
D-37077 Goettingen
GPG Key ID = A46BEE1A
On Tue, 23 Sep 2008, Tommi Kajander wrote:
Hi,
Could anyone give a more or less exact recipee for preparing a derivate
for
lysozyme or proteinase K?
like what used, concentration and soak time. we'd need some data sets
for a
lab course only thing that
really worked so far was lysozyem KI SIRAS (which however doesnt seem to
be
good enough for demonstration
purposes, so we need more)
Thanks very much!
best,
Tommi
Tommi Kajander, Ph.D.
Structural Biology and Biophysics
Institute of Biotechnology
University of Helsinki
Viikinkaari 1
(P.O. Box 65)
00014 Helsinki
Finland
p. +358-9-19158903
[log in to unmask]
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