Domain-specific N-glycosylation of the membrane glycoprotein dipeptidylpeptidase IV (CD26) influences its subcellular trafficking, biological stability, enzyme activity and protein folding.

Dipeptidyl peptidase IV (DPPIV, CD26) is an N-glycosylated type II plasma membrane protein. The primary structure of rat wild-type DPPIV contains eight potential N-glycosylation sites. To investigate the role of N-glycosylation in the function of DPPIV, three of its asparagine residues were separately converted to glutamine by site-directed mutagenesis. The resulting N-glycosylation mutants of rat DPPIV were studied in stable transfected Chinese hamster ovary cells. All three N-glycosylation mutants to DPPIV showed a reduced half-life, as well as differing degrees of inhibition of the processing of their N-glycans. Mutation of the first (Asn83→Gln) or eighth (Asn686→Gln) N-glycoslation site had only a small effect on its enzymatic activity, cell-surface expression and direct formation, whereas the mutation of the sixth N-glycosylation site (Asn319→Gln) abolished the enzymatic activity, eliminated cell-surface expression and prevented the dimerization of the DPPIV protein. The mutant [Glb319]DPPIV is retained in the cytoplasm and its degration was drastically increased. Our data suggest that the N-glycosylation at Ash319 is involved in protein trafficking and correct protein folding. [ABSTRACT FROM AUTHOR]