Transient, lectin-like association of calreticulin with folding intermediates of cellular and viral glycoproteins.
The soluble, calcium-binding protein calreticulin shares high sequence homology with calnexin, a transmembrane chaperone of glycoprotein folding. Our experiments demonstrated that calreticulin, like calnexin, associated transiently with numerous newly synthesized proteins in the endoplasmic reticulum. The population of proteins that bound to calreticulin was partially overlapping with those that bound to calnexin. Hemagglutinin (HA) of influenza virus was shown to associate with both calreticulin and calnexin. Using HA as a model substrate, it was found that both calreticulin- and calnexin-bound HA corresponded primarily to incompletely disulfide-bonded folding intermediates and conformationally trapped forms. Binding of all substrates was oligosaccharide-dependent and required the trimming of glucose residues from asparagine-linked core glycans by glucosidases I and II. In vitro, alpha-mannosidase digestion of calreticulin-bound HA indicated that calreticulin was specific for monoglucosylated glycans. Thus, calreticulin appeared to be a lectin with similar oligosaccharide specificity as its membrane-bound homologue, calnexin. Both are therefore likely to play an important role in glycoprotein maturation and quality control in the endoplasmic reticulum.
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
J R
JR
Peterson
A
A
Ora
P N
PN
Van
A
A
Helenius
D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037281:Calnexin; D037282:Calreticulin; D002240:Carbohydrate Sequence; D006224:Cricetinae; D003553:Cystine / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D011817:Rabbits; D012261:Ribonucleoproteins / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
D002135:Calcium-Binding Proteins; D037282:Calreticulin; C028391:G protein, vesicular stomatitis virus; D006023:Glycoproteins; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D037102:Lectins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011134:Polysaccharides; D012261:Ribonucleoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D037281:Calnexin; D003553:Cystine