Proteasome inhibition compromises direct retention of cytochrome P450 2C2 in the endoplasmic reticulum

Elzbieta Szczesna-Skorupa, Byron Kemper

Research output: Contribution to journalArticlepeer-review

Abstract

To determine whether protein degradation plays a role in the endoplasmic reticulum (ER) retention of cytochromes P450, the effects of proteasomal inhibitors on the expression and distribution of green fluorescent protein chimeras of CYP2C2 and related proteins was examined. In transfected cells, expression levels of chimeras of full-length CYP2C2 and its cytosolic domain, but not its N-terminal transmembrane sequence, were increased by proteasomal inhibition. Redistribution of all three chimeras from the reticular ER into a perinuclear compartment and, in a subset of cells, also to the cell surface was observed after proteasomal inhibition. Redistribution was blocked by the microtubular inhibitor, nocodazole, suggesting that redistribution to the cell surface followed the conventional vesicular transport pathway. Similar redistributions were detected for BAP31, a CYP2C2 binding chaperone; CYP2E1 and CYP3A4, which are also degraded by the proteasomal pathway; and for cytochrome P450 reductase, which does not undergo proteasomal degradation; but not for the ER membrane proteins, sec61 and calnexin. Redistribution does not result from saturation of an ER retention "receptor" since in some cases protein levels were unaffected. Proteasomal inhibition may, therefore, alter ER retention by affecting a protein critical for ER retention, either directly, or indirectly by affecting the composition of the ER membranes.

Original languageEnglish (US)
Pages (from-to)3221-3231
Number of pages11
JournalExperimental Cell Research
Volume314
Issue number17
DOIs
StatePublished - Oct 15 2008

Keywords

  • BAP31
  • Calnexin
  • Cytochrome P450
  • ER-associated degradation (ERAD)
  • Endoplasmic reticulum
  • Intracellular trafficking
  • MG132
  • Proteasome
  • sec61

ASJC Scopus subject areas

  • Cell Biology

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