renaturation of enzymes
dmcbtch at ucl.ac.uk
Thu Jul 20 11:28:06 EST 1995
In article <Pine.SUN.3.91.950720081353.27009A-100000 at verdi>
hroychow at NMSU.EDU (Hiranya Roychowdhury) writes:
> Somebody asked about a protein that can be renatured with full activity.
> As a molecular biologist, the first one that comes to my mind (and I am
> sure most live in dread of this one) is the Omnipresent and Omnipotent
> 'RNase'. Although I have not worked with its biophysical or biochemical
> properties, it is known to renature with most of its activity from almost
> every physical treatment, including the conditions used in autoclaves.
> Only a combination of chemical and physical treatments can effectively rid
> surfaces of this protein and/or its ability to renature.
> Hiranya S. Roychowdhury
> Plant Genetic Engineering Lab.
> Box 3GL, NM State Univ.
> Las Cruces, NM 88003
> Phone: (505) 646-5785
> hroychow at nmsu.edu
I was a little flippant the first time I answered the question with
RNase A, but I don't think there is a better studied enzyme as far as
renaturation goes. It is the classic model for protein folding. When I
was a graduate student, I purified the ATPase "core" of the hsp70
family and was looking at peptide stimulated ATPase activity of the
family. This was in the stone age of peptide sythesis (1980's-pre ABI)
and I had the departmental shop build a little machine to automate the
tedious process abit. The Baldwin lab across the hall ended up using
the machine more than I did to make all these RNase peptides, which
they were studying by circular dichroism and NMR. There were some good
natured debates about what my feeble ATPase was up to, given that their
peptides would just zip up into wonderful native conformations if you
resuspended them in water or some other gloriously complex biochemical
manipulation. If you look up some references from Robert Baldwin's lab,
it will lead you on a trail about anything you could possibly want to
know about RNase A.
MRC Laboratory for Molecular Cell Biology
University College London
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