Biogenesis of plasma membrane glycoproteins. Purification and properties of two rat liver plasma membrane glycoproteins.
As a preliminary to a study of the biogenesis of individual plasma membrane glycoproteins, the marker enzyme nucleotide pyrophosphatase (NPPase) and a major rat liver plasma membrane sialoprotein, subsequently found to be identical with the enzyme dipeptidyl peptidase IV (DPP IV), were purified 10,000- and 2,000-fold, respectively, from rat liver. Both were amphipathic proteins which formed defined micellar complexes with detergents and aggregated in their absence. Gel filtration, sucrose density gradient centrifugation, and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate showed the Triton X-100 complex of NPPase to contain a single 150,000-dalton peptide, while that of DPP IV was composed of two 120,000-dalton subunits; each complex also contained about 150,000-dalton Triton X-100. Trypsin cleaved the detergent complexes with release of major hydrophilic fragments which no longer bound detergent micelles; the accompanying change in peptide size was small for NPPase and undetectable for DPP IV, which also retained the dimer structure of its native form. DPP IV was the only major glycoprotein in rat liver plasma membrane which bound strongly to wheat germ agglutinin. Monospecific rabbit antibodies against NPPase and DPP IV precipitated the antigens without affecting their enzymatic activities.
D000469:Alkaline Phosphatase / Q000302:isolation & purification; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002462:Cell Membrane / Q000201:enzymology*; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000302:isolation & purification* / Q000378:metabolism; D010450:Endopeptidases / Q000302:isolation & purification*; D006023:Glycoproteins / Q000302:isolation & purification*; D007118:Immunoassay; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000302:isolation & purification*; D008970:Molecular Weight; D009711:Nucleotides / Q000302:isolation & purification / Q000378:metabolism; D011487:Protein Conformation; D011755:Pyrophosphatases / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D012795:Sialoglycoproteins / Q000302:isolation & purification*
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D009711:Nucleotides; D012795:Sialoglycoproteins; D000469:Alkaline Phosphatase; D010450:Endopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D011755:Pyrophosphatases; C021899:nucleotide pyrophosphatase