Quality control of transmembrane domain assembly in the tetraspanin CD82.
Retention of misfolded proteins in the endoplasmic reticulum (ER) is a primary mechanism of quality control. To discover whether quality control can monitor assembly inside the hydrophobic ER membrane, we characterized the folding and transport of the tetraspanin glycoprotein CD82. Truncated forms of CD82 that are missing one or more transmembrane segments remain in the ER. A construct (TM 2-4) that is missing the first transmembrane segment remains in the ER, even though its extracellular domain, which is facing the ER lumen, has folded to the native structure. Transport to the cell surface is restored by co-expressing the missing segment (TM 1) as a separate polypeptide. Prior to leaving the ER, CD82 transiently associates with the membrane-bound chaperone calnexin but not with its soluble homolog calreticulin. TM 2-4, in contrast, remains in a prolonged interaction with calnexin that is partially reversed by co-expressing TM 1. These findings establish a simple system to study transmembrane domain assembly, show that ER quality control can directly monitor assembly inside the lipid bilayer and suggest that calnexin may play a role in this process.
Cannon
K S
KS
Section of Immunobiology, Yale University School of Medicine and Howard Hughes Medical Institute, 310 Cedar Street, New Haven, CT 06510, USA.
Cresswell
P
P
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001665:Binding Sites; D001692:Biological Transport; D016466:CHO Cells; D019556:COS Cells; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D037282:Calreticulin; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006031:Glycosylation; D010084:Oxidation-Reduction; D017510:Protein Folding; D012261:Ribonucleoproteins / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D015703:Antigens, CD; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D012261:Ribonucleoproteins; D037281:Calnexin; D014357:Trypsin