"de novo" enzyme identification
RH01358@rh.dk
Thu, 13 Aug 1998 16:24:00 DFT
Dear friends,
I'm starting on a project in a new (to me) ballpark and look for
references (method chapters/books) to give me some genereal starting
guidelines.
The task is to characterize and eventually identify a potentially
new protelytical enzyme involved in post-translational processing of
regulatory peptides. The assay will comprise incubation with the
substrate and detection by RIA of the new exposed terminus. Like other
enzymes involved in post-translational processing it is probably
present in minute amounts. The substrate and products are endogenous
to the cells, so in order to decrease background the enzyme needs
initially to be separated from the lower molecular weight peptides.
Well, these are details which I should maybe not bother you with.
There are many assay conditions and initial purification procedures
which need to be optimized. I have of course a number of ideas, but it
would be convenient to have some general procedure to step along in
the beginning, checklists etc. Are there any publications around that
you could recommend?
Thanks a lot in advance.
best regards
Anders H. Johnsen
Department of Clinical Biochemistry
The National University Hospital
KB3014, Rigshospitalet
Blegdamsvej 9
DK-2100 Copenhagen, DENMARK
phone: (+45) 3545 3007
fax: (+45) 3545 4640
e-mail: johnsen@rh.dk