The p88 molecular chaperone is identical to the endoplasmic reticulum membrane protein, calnexin.
We previously described a novel molecular chaperone (designated p88) that participates in the assembly of murine class I histocompatibility molecules (Degen, E., and Williams, D. B. (1991) J. Cell Biol. 112, 1099-1115). Our findings suggest that p88 may either promote proper assembly of class I molecules or retain them, probably within the endoplasmic reticulum (ER), until assembly of the ternary complex of heavy chain, beta 2-microglobulin, and peptide ligand is complete. In this report, we compare p88 to calnexin, a calcium-binding 90-kDa phosphoprotein of the ER membrane (Wada, I., Rindress, D., Cameron, P. H., Ou, W.-J., Doherty, J.-J., II, Louvard, D., Bell, A.W., Dignard, D., Thomas, D. Y., and Bergeron, J. J. M. (1991) J. Biol. Chem. 266, 19599-19610). We show that p88 and calnexin share antigenic epitopes defined by a polyclonal anti-calnexin antiserum. Furthermore, both proteins were immunoprecipitated in association with an intracellularly retained variant of the class I H-2Kb molecule. Since p88 and calnexin were also indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, were resistant to digestion with endoglycosidase H, and exhibited virtually identical patterns of peptide fragments following digestion with either V8 protease or trypsin, we conclude that p88 and calnexin represent the same protein. The identification of the p88 chaperone as a phosphorylated, calcium-binding protein of the ER membrane suggests possible means whereby its interaction with class I molecules may be regulated.
Department of Biochemistry, University of Toronto, Ontario, Canada.
N
N
Ahluwalia
J J
JJ
Bergeron
I
I
Wada
E
E
Degen
D B
DB
Williams
D000818:Animals; D002135:Calcium-Binding Proteins / Q000737:chemistry*; D037281:Calnexin; D018833:Chaperonins; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D007106:Immune Sera; D008565:Membrane Proteins / Q000737:chemistry*; D051379:Mice; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000737:chemistry*; D011233:Precipitin Tests; D011506:Proteins / Q000737:chemistry*; D014407:Tumor Cells, Cultured
D002135:Calcium-Binding Proteins; D007106:Immune Sera; D008565:Membrane Proteins; D010750:Phosphoproteins; D011506:Proteins; D037281:Calnexin; D018833:Chaperonins