Protein folding within and protein transport into mammalian microsomes are differentially affected by photoaffinity labeling of microsomes with 8-azido-ATP.
Transport of presecretory proteins into mammalian microsomes involves a microsomal protein which is sensitive to photoaffinity labeling with 8-azido-ATP. Typically, protein folding within the lumen of the endoplasmic reticulum of mammalian cells depends on ATP and the member of the Hsp70 protein family, BiP. Here we addressed the question of whether protein transport into and folding within microsomes are differentially affected by photoaffinity labeling of microsomes with 8-azido-ATP. Folding of heterodimeric luciferase to the native state was more azido-ATP-sensitive compared to transport of the precursors of the two subunits. Therefore, we conclude that the microsomal protein which is responsible for the ATP-dependence of protein folding in the endoplasmic reticulum is sensitive to photoaffinity labeling with 8-azido-ATP and that this microsomal protein is distinct from the microsomal ATP-binding protein which is involved in protein transport.
Brunke
M
M
Universität des Saarlandes, Homburg, Germany.
Tyedmers
J
J
Zimmermann
R
R
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000345:Affinity Labels; D000818:Animals; D001386:Azides / Q000378:metabolism*; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D002474:Cell-Free System; D004285:Dogs; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism; D017510:Protein Folding; D011498:Protein Precursors / Q000378:metabolism; D011817:Rabbits
D000345:Affinity Labels; D001386:Azides; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011498:Protein Precursors; C017040:8-azidoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D008156:Luciferases