Dear Gett,
Maybe you want to start by rephrasing the original question.
What you need to know before anything else, is which E2 works together
with your new E3, at least in vitro.
Crystallizing with a random E2 sounds like a very bad idea - even if
that works, why would it be interesting?
Needles to say that this is a very difficult project, unless you are a
lab with considerable expertise in E2/E3 interactions ...
(don't we all love GMAIL addresses for ccp4bb?)
A.
On May 24, 2011, at 15:48, manjeet mukherjee wrote:
Hi all,
I am working on a newly identified E3 ubiquitin ligase (RING type).
I am interested to co-crystallize it with E2 enzyme. Among the
papers reported so far, which are just a few, people have used a
mixture of E2s including UbcH1, UbcH5 and UbcH6 for the
ubiquitination assays. However, there is no clear evidence on E2
enzyme functions with this E3 ubiquitin ligase in vivo.
Can someone suggest how can I choose one particular E2 (among UbcH1/
UbcH5/UbcH6/UncH7...) for co-crystallization studies. It should be
noted that all these E2 enzymes have a similar fold, so will it make
sense to just randomly choose any one of these E2 enzymes and get
started.
Also, if someone with experience in this area suggest weather a Ub-
E2-E3 complex is stable enough and suitable for ternary complexes.
Thanks in advance.
-Geet
P please don't print this e-mail unless you really need to
Anastassis (Tassos) Perrakis, Principal Investigator / Staff Member
Department of Biochemistry (B8)
Netherlands Cancer Institute,
Dept. B8, 1066 CX Amsterdam, The Netherlands
Tel: +31 20 512 1951 Fax: +31 20 512 1954 Mobile / SMS: +31 6 28 597791
