EP24.15 Is Associated with Lipid Rafts

Nathaniel Jeske, Marc J. Glucksman, James Roberts

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Metalloendopeptidase EC 3.4.24.15 (EP24.15, thimet oligopeptidase) is a neuropeptide-metabolizing peptidase expressed throughout the body, but primarily in the brain, gonads, and pituitary. For EP24.15 to have its greatest effect upon peptides in the periphery, it must be targeted and released into the extracellular space. Western blot analysis of fractions taken from discontinuous sucrose density gradients carried out on crude plasma membrane fractions from AtT-20 cells reveals colocalization of EP24.15 and flotillin-1, a known lipid raft marker. Further analysis revealed that an intracellular membrane marker and non-lipid raft, plasma membrane marker, failed to colocalize, supporting EP24.15/lipid raft association. Furthermore, EP24.15 immunoreactivity in lipid raft fractions generated from cells treated with methyl β-cyclodextrin (MβCD) was greatly reduced. Finally, treatment with MPβD resulted in the accumulation of EP24.15 in the media of drug-treated cells over vehicle-treated cells, suggesting that a large percentage of EP24.15 associating with lipid rafts resides on the extracellular surface of the plasma membrane. With this exofacial localization, EP24.15 could have ample access to neuropeptides not only in the immediate microenvironment, but the ability to degrade or modify peptides bound for receptor interaction.

Original languageEnglish (US)
Pages (from-to)468-473
Number of pages6
JournalJournal of Neuroscience Research
Volume74
Issue number3
DOIs
StatePublished - Nov 1 2003

Fingerprint

thimet oligopeptidase
Lipids
Cell Membrane
Neuropeptides
Metalloendopeptidases
Intracellular Membranes
Peptide Receptors
Cyclodextrins
Extracellular Space
Gonads
Sucrose
Peptide Hydrolases
Western Blotting
Peptides
Brain
Pharmaceutical Preparations

Keywords

  • EP24.15
  • Lipid rafts
  • Metalloendopeptidase

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

EP24.15 Is Associated with Lipid Rafts. / Jeske, Nathaniel; Glucksman, Marc J.; Roberts, James.

In: Journal of Neuroscience Research, Vol. 74, No. 3, 01.11.2003, p. 468-473.

Research output: Contribution to journalArticle

Jeske, Nathaniel ; Glucksman, Marc J. ; Roberts, James. / EP24.15 Is Associated with Lipid Rafts. In: Journal of Neuroscience Research. 2003 ; Vol. 74, No. 3. pp. 468-473.
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