Re: [ccp4bb] protease cleavage sites

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: Re: : MTZ to SHELX question
From: Eleanor Dodson ccp4 {- at -} YSBL {- dot -} YORK {- dot -} AC {- dot -} UK
Date: 2007-03-05

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Subject: Re: protease cleavage sites
From: Cynthia Kinsland clk10 {- at -} CORNELL {- dot -} EDU
Date: 2007-03-05

Subject: Re: protease cleavage sites
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-03-05

Rene,

There are many proteases that are suitable for digestion of an N-terminal
tag. Interestingly, there are NOT many (any?) proteases suitable for
digestion of thr C-terminal tag (except for the dual-enzyme His-tag digest
system that sometimes work and sometimes does not).

I personally like TVMV, TEV, and Thrombin. With the first two enzymes, you
have no worries whatsoever about non-specific digestion, whereas Thrombin
is pretty gentle but has been reported to eat things up sometimes.

Cheers,

Artem

> 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
> ===============================================
>
>
>
>

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: : MTZ to SHELX question
From: Eleanor Dodson ccp4 {- at -} YSBL {- dot -} YORK {- dot -} AC {- dot -} UK
Date: 2007-03-05

Next message:
Subject: Re: protease cleavage sites
From: Cynthia Kinsland clk10 {- at -} CORNELL {- dot -} EDU
Date: 2007-03-05