Jenny
have a look at our REFOLD database: http://refold.med.monash.edu.au
you can find several refolding protocols in there for immunoglobulin folds (you can blast
your sequence against the refold database)
cheers
ashley
--
Ashley Buckle Ph.D
NHMRC Senior Research Fellow
The Department of Biochemistry and Molecular Biology
School of Biomedical Sciences, Faculty of Medicine &
Victorian Bioinformatics Consortium (VBC)
Monash University, Clayton, Vic 3800
Australia
http://www.med.monash.edu.au/biochem/staff/abuckle.html
iChat/AIM: blindcaptaincat
skype: ashley.buckle
Tel: (613) 9902 0269 (office)
Tel: (613) 9905 1653 (lab)
Fax : (613) 9905 4699
----- Original Message -----
From: Jenny <[log in to unmask]>
Date: Thursday, July 10, 2008 11:00 am
Subject: [ccp4bb] how to promote folding for an unfolded protein
To: [log in to unmask]
> Dear CCP4 community,
>
> Sorry for this off-topic protein purification problem.I'm trying to
> purifyan immunoglobulin-like beta sheet protein with a c-terminus
> HIS construct.
> The protein expressed both in the supernat and pellet ( majority ). I
> purified the supernat and after run gel filtration, it's in the
> void volume.
> I also tried to purify from the pellet,and do dialysis refolding (
> with and
> without L-arg ), after overnight dialysis, I run the gel filtration
> column,the apparent molecular weight looks like dimer/trimer. But
> when I did a CD
> scan of the protein, it's showing an unfolded protein profile.I was
> wondering if there is anyway to promote folding,or if anyway that
> can make
> some mutations to make it foldable.Any input would be useful.
>
> Thanks a lot.
>
> Jenny
>
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