[ccp4bb]: Summary MIR-test case

[Hemant Yennawar <hpy1@psu.edu>]

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Hello everyone,
    I thank everyone who had suggestions. I will copy all the e-mails here 
(some were directly submitted to the CCP4BB) for completeness:

Michael Sawaya <sawaya@mbi.ucla.edu> wrote:
Proteinase K would be an excellent choice. GdCl3, SmCl3,
and p-hydroxymercuribenzoic acid make high quality derivatives.
We used proteinase K and derivatives in a structural molecular
biology lab course recently given at UCLA.
Instructions for crystallization, heavy atom preparation, and where
to buy proteinase K can be found at:
http://www.doe-mbi.ucla.edu/~sawaya/m230d/Crystallization/crystallization.html


David Briggs <d.briggs@mail.cryst.bbk.ac.uk> whote:
I think that the good old favorite Lysozyme has been solved using a K2 UO2
F5 derivative and a mercury derivative as well (can't remember which)
The details are in Blundell and Johnson...

"Fred. Vellieux" <vellieux@ibs.fr> wrote:
If you are interested, I can provide you with a copy of the "Introduction
to PX" course notes that I have. I gave this course several times over the
years, the introduction is a practical (hands-on) approach. It uses
lysozyme, and a mercury derivative (that was not used during
structure-determination by David Phillips and colleagues due to slight
non-isomorphism). "Standard case" I would say.
Please let me know if you are interested. Some of the notes should be
updated to take advantage of the new technologies (area detectors ...).

Gabby Rudenko <rudenko@chop.swmed.edu> wrote:
>I'm sure many people will mention lyzozyme and as derivative
>    lyzozyme + PCMBS (para-chloro-mercuric benzene sulfonic acid). You
>    co-cystallize the latter actually; The patterson is very nice.
>    This example is often used in workshops.
>
>    E-mail me a fax. nr. if you need more details.

Jim Pflugrath <jwp@RigakuMSC.com> wrote:
In the CSHL course we have problems getting hen egg white lysozyme
derivatives. We have used lysozyme, thaumatin (with Hg), and glucose
isomerase (with Hg). All without much success. We now use lysyzome
crystallized in the presence of NaAu(CN)2 which gives orthorhombic
crystals so we know the Au is incorporated. See Yang & Pflugrath (2001)
Acta Cryst. D57, 1480-1490. While those crystals are great for SAD or
MAD, you have no native crystal for MIR. As you'll read in the paper, it
seems that the orthorhombic crystals lose Au as they get older, so you can
get a native in due time. Maybe you can work out a way to get rid of the
Au by an EDTA or some other soak.
Also, pCMBS apparently co-crystallizes with lysozyme, but the unit cell
changes slightly, the substitution is not great and the phases are poor.


Thank you all,

Hemant

Hemant Yennawar Ph.D.           Phone   814 865 8383
8A Althouse Laboratory,         Fax     814 863 7024
University Park, PA 16802