Dissociation of complexes between 70 kDa stress proteins and presecretory proteins is facilitated by a cytosolic factor.
Members of the 70 kDa stress protein family were shown previously to facilitate the posttranslational translocation of presecretory proteins into the endoplasmic reticulum and protein precursors into mitochondria. To identify proteins that interact with 70 kDa stress proteins during the early steps of posttranslational translocation, polyclonal antibodies were raised against purified yeast cytosolic stress proteins. They were used to immunoprecipitate complexes between 70 kDa stress proteins and a radiolabeled presecretory protein, prepro-alpha-factor, that was translated in vitro. Complexes between prepro-alpha-factor and 70 kDa stress proteins were stable, but could be dissociated in the presence of ATP and crude cytosolic extracts from yeast. These results are consistent with the idea that 70 kDa stress proteins act as molecular chaperones in translocation by binding to precursor proteins before or during their passage across membranes.
Chirico
W J
WJ
Department of Anatomy and Cell Biology, State University of New York-Health Science Center, Brooklyn 11203.
D000906:Antibodies; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D005033:Ethylmaleimide / Q000494:pharmacology; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000302:isolation & purification / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins
D000906:Antibodies; D005656:Fungal Proteins; D006360:Heat-Shock Proteins; C048632:MF(ALPHA)1 protein, S cerevisiae; D011498:Protein Precursors; D029701:Saccharomyces cerevisiae Proteins; D010450:Endopeptidases; D005033:Ethylmaleimide