PROSTAGLANDIN 19- &20-HYDROXYLATION BY CYTOCHROME P-450

  • Masters, Bettie S (PI)

Project: Research project

Project Details

Description

For the past 10 years, the Principal Investigator's laboratory has been
interested in the hydroxylation of medium-chain fatty acids and
prostaglandins by kidney, liver, and lung microsomal preparations and
purified preparations of cytochromes P-450 derived from these tissues.
Hydroxylation of lauric acid and prostaglandins E1, A1, and F2Alpha occurs
mainly in the Omega- and (Omega-1)-positions leading to the production of
products which can be further oxidized by cytosolic dehydrogenases to
dicarboxylic acids. The formation of high concentrations of
Omega-hydroxylated products of prostaglandins E1 and F2Alpha by lung
microsomes from pregnant rabbits and the occurrence of
(Omega-1)-hydroxyprostaglandin E1 in primate semen at high levels has led
to increased interest in the metabolism of these prostaglandins in
reproductive biology. Experiments are being proposed to: 1) localize the
cytochrome(s) P-450 in lung tissue responsible for the Omega- hydroxylase
activities by fluorescent antibody labelling and/or colloidal gold-labelled
antibody techniques; 2) determine the fate of the Omega-hydroxylated
metabolites of fatty acids and prostaglandins by lung perfusion techniques;
3) test the physiological activities of the Omega-hydroxyprostaglandins vs
the parent compounds; 4) determine the specificity of the
Omega-hydroxylase(s) in purified, reconstituted systems for prostaglandins
and their precursors in the presence and absence of cytochrome b5; 5)
localize the male reproductive organ responsible for the formation of and
determine the precursor(s) of the (Omega-1)-hydroxy PGE1 in male primate
semen; 6) isolate and purify the (Omega-1)-hydroxylation system from the
appropriate organ if localization is possible and tissue is obtainable; 7)
utilization of the antibody to lung microsomal prostaglandin
Omega-hydroxylase cytochrome P-450 (P-450 PG-Omega) to isolate the in vitro
translation products of total RNA isolated from pregnancy- or
progesterone-induced rabbits and isolation of specific mRNAs to obtain a
cDNA probe for partial sequence analysis and comparison to other
cytochromes P-450; and 8) cloning of the specific cDNA sequence of
cytochrome P-450PG-Omega selected by the hybrid arrest technique and
confirmed by the positive hybridization-translation assay. These studies
are designed to determine the metabolism of, mechanism of action of, and
molecular regulation for Omega- and (Omega-1)-hydroxylated prostaglandins
E1 and F2Alpha.
StatusFinished
Effective start/end date6/1/826/30/07

Funding

  • National Institutes of Health: $321,346.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $194,544.00
  • National Institutes of Health: $169,691.00
  • National Institutes of Health
  • National Institutes of Health: $291,132.00
  • National Institutes of Health: $209,845.00
  • National Institutes of Health
  • National Institutes of Health: $212,968.00
  • National Institutes of Health
  • National Institutes of Health: $271,756.00
  • National Institutes of Health
  • National Institutes of Health: $218,891.00
  • National Institutes of Health: $223,616.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $193,653.00
  • National Institutes of Health: $302,783.00

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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