Isolation and Characterization of the Rhesus Monkey Renal 15-Hydroxy Prostaglandin Dehydrogenase
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Authors
Lambert, Jack S.
Issue Date
1975
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Isolation of the Rhesus monkey renal 15-hydroxydehydrogenase
resulted in an approximately 85-fold purification by
using DEAE-cellulose and hydroxylapatite chromatography purification
methods. Disc gel electrophoresis showed several protein bands, indicating
the partial purity of the enzyme. Assay studies indicated that the
specific cofactor for the enzyme was NAD+; NADP+ was not a cofactor.
The best substrates were PGE1, PGA1, PGE2 and PGA2, as indicated by
Km of 8.0 ~M, 7.5 ~M, 7.8 ~M, and 9.26 ~M, respectively. The prostaglandin
analogs 17-~-PGE1 and ~4-PGE1 had Km of 12.2 ~M and 27.4 ~M,
respectively; with the PGF1a and PGF2a being substrates to a lesser
degree, 31.25 ~M and 62.5 ~M. The following compounds were poor substrates
or not substrates at all: PGB1, PGB2, PGD1' 5-oxa PGF1a, l5(R) PGE2'
16,16-dimethyl PGFla, and 15-methy1 PGE2. Of the inhibitors tested,
ethacrynic acid, furosamide, and prostanoic acid exhibited inhibition,
whereas angiotensin II, eledoisin, and bradykinin did not. The results
suggest a successful method for enzyme purification, show the specificity
of the enzyme, and help elucidate the mode of action of certain known
inhibitors of renal function.
Description
v, 28 p.
Citation
Publisher
Kalamazoo College
License
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