The role of ERp57 in disulfide bond formation during the assembly of majorhistocompatibility complex class I in a synchronized semipermeabilized cell translation system
Mr. Farmery et al., The role of ERp57 in disulfide bond formation during the assembly of majorhistocompatibility complex class I in a synchronized semipermeabilized cell translation system, J BIOL CHEM, 275(20), 2000, pp. 14933-14938
We have established a semipermeabilized cell system that reproduces the fol
ding and assembly of a major histocompatibility complex (MHC) class I compl
ex as it would occur in the intact cell. The translation of the MHC class I
heavy chain (HLA-B27) in this system was synchronized allowing the folding
and assembly of polypeptide chains synthesized within a short time frame t
o be analyzed. This has enabled us to dissect the time course of interactio
n of both disulfide and nondisulfide-bonded heavy chain with various molecu
lar chaperones during its assembly in a functionally intact endoplasmic ret
iculum, The results demonstrate that unassembled, nondisulfide-bonded forms
of heavy chain interact initially with calnexin, A later and more prolonge
d interaction of calreticulin, specifically with assembled, disulfide-bonde
d heavy chain, highlights distinct differences in the roles of these two pr
oteins in the assembly of MHC class I molecules. We also demonstrate that t
he thiol-dependent reductase ERp57 initially interacts with nondisulfide-bo
nded heavy chain, but this rapidly becomes disulfide-bonded and indicates t
hat heavy chain folding occurs during its interaction with ERp57, In additi
on, we also confirm a direct interaction between MHC class I heavy chain an
d tapasin, emphasizing the role that this protein plays in the later stages
of MHC class I assembly.