Specific isoforms of the resident endoplasmic reticulum protein glucosidase II associate with the CD45 protein-tyrosine phosphatase via a lectin-like interaction.
We have previously demonstrated that CD45 physically associates with the endoplasmic reticulum processing enzyme glucosidase II (GII). GII consists of the catalytic alpha-chain and an associated beta-chain. To gain insight into the basis of the association between CD45 and GII, we examined the biochemical requirements for the interaction. We show that the alpha-subunit is essential for the interaction. Interestingly, only a higher molecular weight form of GIIalpha is capable of associating with CD45 in a competitive situation where multiple GIIalpha isoforms are expressed. Further, transfection studies demonstrate that only isoforms containing the alternatively spliced sequence Box A1 are capable of binding CD45, although all isoforms are catalytically active. The interaction between CD45 and GII is dependent on the active site of GII, is mediated through the carbohydrate on CD45, and can be inhibited with mannose. Taken together, these results suggest that GIIalpha acts as a lectin and binds to CD45 in an exon-dependent manner. This lectin activity of GII may be a novel mechanism for the regulation of CD45 biology and play a role in immune function, possibly by regulating CD45 glycosylation.
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, T6G 2S2 Alberta, Canada.
T A
TA
Baldwin
M
M
Gogela-Spehar
H L
HL
Ostergaard
D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001667:Binding, Competitive; D002384:Catalysis; D020134:Catalytic Domain; D004721:Endoplasmic Reticulum / Q000201:enzymology* / Q000378:metabolism; D005091:Exons / Q000235:genetics; D006031:Glycosylation; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D037102:Lectins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D008358:Mannose / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D011485:Protein Binding / Q000187:drug effects; D021122:Protein Subunits; D014162:Transfection; D014407:Tumor Cells, Cultured; D000520:alpha-Glucosidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D007527:Isoenzymes; D037102:Lectins; D021122:Protein Subunits; D017493:Leukocyte Common Antigens; C034251:4-nitrophenyl-alpha-glucosidase; D000520:alpha-Glucosidases; D008358:Mannose