Regulation of cell-surface major histocompatibility complex class I expression by the endopeptidase EC3.4.24.15 (thimet oligopeptidase)

Sandra I. Kim, Amanda Pabon, Todd Swanson, Marc J. Glucksman

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Endopeptidase EP24.15 (EC 3.4.24.15; thimet oligopeptidase), traditionally classified as a neuropeptide-processing enzyme, degrades well-known MHC I (major histocompatibility complex class I) peptides in cell extracts. In the present study, we determine the contribution of EP24.15 in vivo to the surface expression of MHCI on intact cells. CTLs (cytotoxic T-lymphocytes) recognize a vast array of peptides presented in the context of MHC I cell-surface molecules. Stable retroviral overexpression of EP24.15 induces a dramatic, long-term inhibition of surface MHC I. In contrast, overexpression of a mutant EP24.15, which is expressed, but is enzymically inactive, does not affect the surface MHC I expression level. We observed no difference in the effect of EP24.15 on the expression of different classes of MHC I. IFN-γ (interferon-γ) treatment re-established MHC I expression on these EP24.15-overexpressing cells, and also induced EP24.15 cytosolic protein expression and enzyme activity. To our knowledge, this is the first demonstration of cytokine-induced EP24.15 expression and activity. Conversely, stable retroviral silencing of endogenous EP24.15 by RNA interference induced a striking, long-term increase in surface MHC I. Subcellular fractionation and enzyme-activity experiments localized the vast majority of EP24.15 protein expression and function to the cytosol. Therefore we introduce a novel function of the cytosolic form of EP24.15. EP24.15 activity in the extracellular space is significant for neuropeptide processing, and in the present paper, we demonstrate that EP24.15 activity in the cytosol may be significant for regulation of MHC I cell-surface expression.

Original languageEnglish (US)
Pages (from-to)111-120
Number of pages10
JournalBiochemical Journal
Volume375
Issue number1
DOIs
StatePublished - Oct 1 2003
Externally publishedYes

Fingerprint

thimet oligopeptidase
Endopeptidases
Major Histocompatibility Complex
Cells
Neuropeptides
Cytosol
Enzymes
Enzyme activity
Peptides
Extracellular Space
Cytotoxic T-Lymphocytes
RNA Interference
Cell Extracts
Interferons
Proteins
T-cells
Fractionation
Cytokines
Processing
Demonstrations

Keywords

  • Cytosol
  • Endopeptidase 24.15 (EP24.15)
  • Major histocompatibility complex class I (MHC I)
  • Metallopeptidase
  • Neuropeptide
  • Thimet oligopeptidase (TOP)

ASJC Scopus subject areas

  • Biochemistry

Cite this

Regulation of cell-surface major histocompatibility complex class I expression by the endopeptidase EC3.4.24.15 (thimet oligopeptidase). / Kim, Sandra I.; Pabon, Amanda; Swanson, Todd; Glucksman, Marc J.

In: Biochemical Journal, Vol. 375, No. 1, 01.10.2003, p. 111-120.

Research output: Contribution to journalArticle

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