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Re: [ccp4bb] protease cleavage sites
 

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CCP4bb <-- 2007 <-- March 2007 <-- 05 March 2007
Previous message:
Subject: AW: MTZ format conversion
From: Herman {- dot -} Schreuder {- at -} SANOFI-AVENTIS {- dot -} COM Herman {- dot -} Schreuder {- at -} SANOFI-AVENTIS {- dot -} COM
Date: 2007-03-05 		Next message:
Subject: Re: MTZ to Shel-X?
From: "George M {- dot -} Sheldrick" gsheldr {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2007-03-05


Subject: Re: protease cleavage sites
From: price {- at -} UCHICAGO {- dot -} EDU price {- at -} UCHICAGO {- dot -} EDU
Date: 2007-03-05
TeV is great but it isn't always a magic bullet for producing native
N-termini: we've experimentally determined the obvious, that if you
bury part of its recognition site in secondary structure, it cleaves
very very slowly. We tried this on a protein where M1 is cleaved in
vivo and aa#2 is the beginning of a nice helix, but the sequence
itself is compatible with TeV sites. Adding a couple small flexible
residues between the cleavage site and the beginning of the folded
structure did wonders for the cleavage rate.
Phoebe

At 04:01 AM 3/2/2007, Rene Frank wrote:
>Hi,
>
>A non-ccp4 Q. Sorry.
>
>I would like to use a cleavable purification tag at the
>N-terminus/extracellular end of my membrane protein for
>purification. Before I start, I wonder if someone could recommend a
>particular protease site that I can engineer between the tag and my
>protein? How about a proprietary cleavage system such as the
>PreScission protease (GE Healthcare)? I would be grateful to hear
>success and horror stories in this area.
>
>Best wishes,
>
>Rene
>
>================================================
>Dr R.A.W. Frank, PhD
>Royal Commission for the Exhibition of 1851 Research Fellow
>
>Prof Seth Grant Lab / Genes to Cognition
>Wellcome Trust Sanger Institute
>Hinxton
>Cambridge CB10 1SA
>
>Work Tel: 0044 (0)1223 834244 ext. 7318
>Cell No.: 0044 (0)7870 208280
>===============================================
>
>

---------------------------------------------------------------------------------------------------------------------------
Phoebe A. Rice
Assoc. Prof., Dept. of Biochemistry & Molecular Biology
The University of Chicago
phone 773 834 1723
fax 773 702 0439
http://bmb.bsd.uchicago.edu/index.html
http://www.nasa.gov/mission_pages/cassini/multimedia/pia06064.html

CCP4bb navigation

CCP4bb <-- 2007 <-- March 2007 <-- 05 March 2007
Previous message:
Subject: AW: MTZ format conversion
From: Herman {- dot -} Schreuder {- at -} SANOFI-AVENTIS {- dot -} COM Herman {- dot -} Schreuder {- at -} SANOFI-AVENTIS {- dot -} COM
Date: 2007-03-05 		Next message:
Subject: Re: MTZ to Shel-X?
From: "George M {- dot -} Sheldrick" gsheldr {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2007-03-05


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