Efficient folding of firefly luciferase after transport into mammalian microsomes in the absence of luminal chaperones and folding catalysts.
Folding of polypeptides emerging from the protein translocase in the membrane of mammalian microsomes was analyzed after synthesis of corresponding precursor proteins in a mammalian translation system. Firefly luciferase was used as a model protein; the corresponding hybrid precursor contained the preprolactin signal peptide. The rates and efficiencies of folding of luciferase in microsomes were compared with those of folding of luciferase in the cytosol. Furthermore, folding of luciferase in microsomes was compared with that in proteoliposomes, i.e. in the absence of luminal molecular chaperones and folding catalysts. Folding in microsomes was less efficient compared with folding in the cytosol. Folding in the absence of luminal proteins was more efficient compared with folding in their presence and identical to folding in the cytosol. Thus, firefly luciferase emerging from translocase can efficiently fold to its native conformation without chaperoning by any luminal proteins. There may be molecular chaperones present in the microsomal membrane that can efficiently substitute for the cytosolic chaperone machinery comprising Hsp40, Hsp60, and Hsp70 with respect to folding of firefly luciferase.
Tyedmers
J
J
Medizinische Biochemie, Universität des Saarlandes, D-66421 Homburg, Federal Republic of Germany.
Brunke
M
M
Lechte
M
M
Sandholzer
U
U
Dierks
T
T
Schlotterhose
P
P
Schmidt
B
B
Zimmermann
R
R
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001386:Azides / Q000494:pharmacology; D001692:Biological Transport; D002384:Catalysis; D001517:Coleoptera / Q000201:enzymology*; D016572:Cyclosporine / Q000494:pharmacology; D004285:Dogs; D008156:Luciferases / Q000378:metabolism*; D008861:Microsomes / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D017510:Protein Folding; D011510:Proteolipids / Q000378:metabolism; D016559:Tacrolimus / Q000494:pharmacology; D014466:Ultraviolet Rays
D001386:Azides; D018832:Molecular Chaperones; D011510:Proteolipids; C020485:proteoliposomes; C017040:8-azidoadenosine 5'-triphosphate; D016572:Cyclosporine; D000255:Adenosine Triphosphate; D008156:Luciferases; D016559:Tacrolimus