Membrane-bound domain of HMG CoA reductase is required for sterol-enhanced degradation of the enzyme

Cell. 1985 May;41(1):249-58. doi: 10.1016/0092-8674(85)90078-9.

Abstract

3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMG CoA reductase) is a single polypeptide chain with two contiguous domains: a soluble domain (548 amino acids) that catalyzes the rate-controlling step in cholesterol synthesis and a membrane-bound domain (339 amino acids) that anchors the protein to the endoplasmic reticulum (ER). HMG CoA reductase is degraded at least 10-fold more rapidly than other ER proteins; degradation is accelerated in the presence of cholesterol. To understand this controlled degradation, we transfected reductase-deficient Chinese hamster ovary (CHO) cells with a plasmid expression vector containing a reductase cDNA that lacks the segment encoding the membrane domain. The plasmid produced a truncated reductase (37 kd smaller than normal) that was enzymatically active with normal kinetics; most of the truncated enzyme was found in the cytosol. The truncated enzyme was degraded one-fifth as fast as the holoenzyme; degradation was no longer accelerated by sterols. We conclude that the membrane-bound domain of reductase plays a crucial role in the rapid and regulated degradation of this ER protein.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Cholesterol / pharmacology*
  • Cricetinae
  • Cytosol / enzymology
  • DNA, Recombinant
  • Endoplasmic Reticulum / enzymology*
  • Female
  • Hydroxycholesterols / pharmacology*
  • Hydroxymethylglutaryl CoA Reductases / genetics
  • Hydroxymethylglutaryl CoA Reductases / metabolism*
  • Intracellular Membranes / enzymology*
  • Kinetics
  • Lovastatin* / analogs & derivatives*
  • Naphthalenes / pharmacology
  • Ovary
  • Plasmids
  • Transfection

Substances

  • DNA, Recombinant
  • Hydroxycholesterols
  • Naphthalenes
  • mevastatin
  • 25-hydroxycholesterol
  • Cholesterol
  • Lovastatin
  • Hydroxymethylglutaryl CoA Reductases