title: Subcellular distribution of the enzymes degrading thyrotropin releasing hormone and metabolites in rat brain.
abstract: In order to further understand the role of enzymes degrading Thyrotropin Releasing Hormone (TRH, pglu-his-proNH(2)) and metabolites, we studied their subcellular distribution in rat brain. Brain tissue was homogenized in 0.32 M sucrose, tris-HCl 0.01 M pH 7.4 and fractionated by differential and discontinuous gradient centrifugation; [(3)H]pro-TRH was incubated with the various subcellular fractions and the extent of degradation of each metabolite was measured after separation by thin layer chromatography. Several markers were simultaneously measured (lactate dehydrogenase, 5?-nucleotidase and hexosaminidase) to determine the pattern of distribution of the subcellular organelles. The post-proline cleaving enzyme responsible for pglu-his-pro formation and pyroglutamate amino-peptidase (which requires sulphydryl compounds for maximal activity) were found in cytosol but were barely detectable in the soluble component of synaptosomes; pyroglutamate aminopeptidase (dependent on metals) and post-proline dipeptidyl amino peptidase were found on the membranes of synaptosomes; imido peptidase was not enriched in any particular fraction. These data are consistent with the hypothesis that membrane-bound pyroglutamate aminopeptidase is responsible for TRH degradation once released into the synaptic cleft and that the post-proline dipeptidylaminopeptidase may participate in the extracellular catabolism of his-proNH(2) before it cyclizes to his-pro-DKP. They also suggest that post-proline cleaving enzyme and soluble pyroglutamate aminopeptidase may not play an important role in the regulation of TRH levels in nerve endings.
authors.affiliation: Centro de Investigación sobre Ingenieria Genética y Biotecnologia Universidad Nacional Autónoma de México, Apartado Postal 70479, México, D.F., 04510, México.
authors.forename: H
authors.initials: H
authors.lastname: Torres
authors.forename: J L
authors.initials: JL
authors.lastname: Charli
authors.forename: A
authors.initials: A
authors.lastname: González-Noriega
authors.forename: M A
authors.initials: MA
authors.lastname: Vargas
authors.forename: P
authors.initials: P
authors.lastname: Joseph-Bravo