Protein transport by purified yeast Sec complex and Kar2p without membranes

Science. 1997 Aug 15;277(5328):938-41. doi: 10.1126/science.277.5328.938.

Abstract

Posttranslational protein translocation across the endoplasmic reticulum membrane of yeast requires a seven-component transmembrane complex (the Sec complex) in collaboration with the lumenal Kar2 protein (Kar2p). A translocation substrate was initially bound to the cytosolic face of the purified Sec complex in a signal-sequence-dependent but Kar2p- and nucleotide-independent manner. In a subsequent reaction, in which Kar2p interacted with the lumenal face of the Sec complex and hydrolyzed adenosine triphosphate, the substrate moved through a channel formed by the Sec complex and was released at the lumenal end. Movement through the channel occurred in detergent solution in the absence of a lipid bilayer.

Publication types

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

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Biological Transport
  • Cross-Linking Reagents
  • Cytosol / metabolism
  • Detergents
  • Digitonin
  • Endoplasmic Reticulum / metabolism
  • Fungal Proteins / metabolism*
  • HSP70 Heat-Shock Proteins / metabolism*
  • Heat-Shock Proteins*
  • Lipid Bilayers
  • Liposomes / metabolism
  • Membrane Proteins / metabolism*
  • Membrane Transport Proteins*
  • Protein Precursors / metabolism*
  • Protein Sorting Signals / metabolism
  • Proteolipids / metabolism
  • RNA, Transfer / metabolism
  • SEC Translocation Channels
  • Saccharomyces cerevisiae Proteins*
  • Solubility
  • Succinimides

Substances

  • Cross-Linking Reagents
  • Detergents
  • Fungal Proteins
  • HSP70 Heat-Shock Proteins
  • Heat-Shock Proteins
  • KAR2 protein, yeast
  • Lipid Bilayers
  • Liposomes
  • MF(ALPHA)1 protein, S cerevisiae
  • Membrane Proteins
  • Membrane Transport Proteins
  • Protein Precursors
  • Protein Sorting Signals
  • Proteolipids
  • SEC Translocation Channels
  • SEC62 protein, S cerevisiae
  • SEC63 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Succinimides
  • proteoliposomes
  • Adenosine Triphosphate
  • RNA, Transfer
  • Digitonin
  • disuccinimidyl suberate