title: N-linked oligosaccharides affect the enzymatic activity of CD39: diverse interactions between seven N-linked glycosylation sites.
abstract: Rat CD39, a membrane-bound ectonucleoside triphosphate diphosphohydrolase that hydrolyzes extracellular nucleoside tri- and diphosphates, has seven potential N-glycosylation sites at asparagine residues 73, 226, 291, 333, 375, 429, and 458. To determine their roles in the structure and function of CD39, we mutated these sites individually or in combination by replacing asparagine with serine or glutamine and analyzed the surface expression and the enzymatic activity of the mutants. The results indicate that rat CD39 can be glycosylated at all seven sites when expressed in COS7 cells. Glycosylation sites 73 at the N terminus, 333 in the middle, and 429 and 458 at the C terminus were principally required for cell surface appearance of enzymatically active CD39. Whereas deletion of these sites individually had modest effects on surface ATPase activity, some double deletions of these sites had major effects on both surface activity and expression. The importance of these N-glycosylation sites is recognizable in other members of the ectonucleoside triphosphate diphosphohydrolase family.
authors.affiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: James J
authors.initials: JJ
authors.lastname: Wu
authors.forename: Lisa E
authors.initials: LE
authors.lastname: Choi
authors.forename: Guido
authors.initials: G
authors.lastname: Guidotti
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D001081:Apyrase / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D002118:Calcium / Q000494:pharmacology; D002522:Chlorocebus aethiops; D004795:Enzyme Stability; D015971:Gene Expression Regulation, Enzymologic; D006031:Glycosylation; D009154:Mutation / Q000235:genetics; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D051381:Rats
chemical_list: D015703:Antigens, CD; D009844:Oligosaccharides; D001081:Apyrase; C067869:CD39 antigen; D002118:Calcium