Characterization of a new plasma membrane-associated ecto-5'-phosphodiesterase/nucleotide-pyrophosphatase from rat hepatocarcinoma AS-30D cells.
We have identified in plasma membrane fractions isolated from rat hepatocarcinoma AS-30D ascites cells three glycoproteins of 125 kDa, 115 kDa and 105 kDa (gp125, gp115 and gp105) which become adenylylated using ATP as substrate, most readily in the presence of EDTA. The gp115 becomes also phosphorylated. The adenylylation of these tumor glycoproteins was much lower than that of a group of analogous adenylylatable glycoproteins (gp130, gp120-gp110 dimer and gp100) present in normal rat liver plasma membrane. The tumor glycoproteins were reversibly O-adenylylated at threonine residues, as was the case for their normal rat liver counterparts. The tumor gp115, and the gp120-gp110 dimer from normal rat liver were both isolated using either ATP-affinity chromatography and/or AMP-affinity chromatography. The gp120-gp110 dimer from normal rat liver was identified as the plasma cell differentiation antigen-1 (PC-1 protein), an ecto-5' phosphodiesterase/ nucleotide-pyrophosphatase (5'-PDE/NPPase). The gp115 from tumor cells also exhibited Zn2+-stimulated 5'-PDE and NPPase activities in alkaline conditions, although it appears to be distinct from the PC-1 protein. We have determined that the gp115 is an ecto-enzyme that catalyzes the hydrolysis of extracellular ATP, since its adenylylation and phosphorylation were detected in intact cells using extracellularly added [alpha-32P]ATP or [gamma-32P]ATP, respectively, in the absence of any permeabilizing agent.
García-Nieto
R M
RM
Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Cientificas and Universidad Autónoma de Madrid, Spain.
José
E S
ES
Martín-Nieto
J
J
Villalobo
A
A
D000227:Adenine Nucleotides / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002846:Chromatography, Affinity; D011756:Diphosphates / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000201:enzymology* / Q000473:pathology; D008562:Membrane Glycoproteins / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D000227:Adenine Nucleotides; D011756:Diphosphates; D008562:Membrane Glycoproteins; D010727:Phosphoric Diester Hydrolases