87-kDa PARP activity

Silas Bruun Silas at knus.dk
Fri Oct 5 03:58:55 EST 2001











Hi
check this new article. PARP-1 is cleaved in such a way that its catalytic
site is in its p85 cleavage fragment. My guess would be that the cleaved
fragment is still active allthough I have not seen this in any paper. The
full enzyme (according to this article) is activated by DNA damage. So mayby
you could activate your protein in some way, maybe by exposing it to nicked
DNA or what do I know.
Good luck

Silas

     Soldani C, Lazze MC, Bottone MG, Tognon G, Biggiogera M, Pellicciari
CE, Scovassi AI. Related Articles
Poly(adp-ribose) polymerase cleavage during apoptosis: when and where?
Exp Cell Res. 2001 Oct 1;269(2):193-201.
PMID: 11570811 [PubMed - in process]

***************
Silas Bruun
The Protein Laboratory
Institute of Molecular Pathology
University of Copenhagen
Blegdamsvej 3 Bldg. 6.2
2200 Cph N, Denmark
fax: +45 353-60116
"Araluen Freeman" <araluen_f at nospam.geocities.com> wrote in message
news:9pcn7d$esl$1 at possum.melbpc.org.au...
>
> Hi all,
>
> I've been getting some weird results with Western and activity blots of
> poly(ADP-ribose) polymerase (PARP).  I was wondering if someone else has
> worked with PARP also, and might share some of their observations of the
> enzyme.
>
> Simply put, we're detecting the full PARP-1 enzyme at ~120-kDa with the
> C2-10 anti-PARP antibody, as well as the 87-kDa cleavage product (AND a
> 100-kDa band, but that's another story).  However, in the corresponding
> activity blot, we aren't getting one hint of activity from the full
enzyme,
> instead picking up distinct basal activity from the 87-kDa cleavage
product.
>
> What we're baffled by is the lack of activity from the full enzyme.
There's
> a few ideas flying around (there always are), but I thought I'd test the
> waters to see if there is anyone else that might have had their paws on
one
> of the PARPs and have an idea.  One suggestion has been that perhaps the
> ~120-kDa PARP-1 we're detecting is already modified, preventing the in
vitro
> activity, but reports have shown that significant self-modification leads
to
> slower electrophoretic mobilities -- our Western is giving us the same
tidy
> band at ~120-kDa.
>
> Anyways, thanking anyone for their suggestions!  ;-)
>
>  Araluen.
>
>





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