High-performance liquid chromatographic analysis of in vitro central neuropeptide processing

Thomas P. Davis, Alison Culling-Berglund

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Reversed-phase high-performance liquid chromatography (HPLC) was used to study and characterize the in vitro proteolytic processing of β-endorphin by twice-washed membrane homogenates. A high-resolution method, capable of separating over 30 different human β-endorphin-related fragments in a single analysis, was used to study the time course of production of specific, biologically active endorphin fragments by membrane-associated proteases. The results demonstrate that frozen (-37°C), postmortem human and rat brains are viable for processing studies and that metabolism proceeds similarly to that in fresh brain homogenates or slices. Significant differences were noted in the formation rates of putative neuroleptic peptides between sex- and age-matched postmortem brain tissues from controls versus postmortem brain tissues from neuropsychiatric patients or drug-treated animals. These data suggest that using HPLC to characterize neuropeptide processing in human or rat membrane-associated enzyme homogenates is both descriptive and quantitative and offers insight into the central regulation of neuropeptide metabolism.

Original languageEnglish (US)
Pages (from-to)279-292
Number of pages14
JournalJournal of Chromatography A
Volume327
Issue numberC
DOIs
StatePublished - Jun 26 1985

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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