Endogenous hyperphosphorylation in plasma membrane from an ascites hepatocarcinoma cell line.
Plasma-membrane-bound kinases of AS-30D ascites from transplantable rat hepatocarcinoma were shown to extensively catalyze the phosphorylation of plasma membrane proteins and membrane lipids, using [gamma-32P]ATP or [gamma-32P]GTP as a phosphate donor. In contrast, plasma membranes from normal adult rat liver or fast-growing regenerating liver (24 h after partial hepatectomy) produce significantly less activity for protein phosphorylation and little phosphorylation of the lipids. However, neonatal (24 h old) rat liver plasma membrane preparations show levels of phosphorylation of proteins and lipids intermediate between those in the tumor cell line and normal adult plasma membrane preparations. Phosphatidic acid was identified as one of the 32P-labelled lipids in the tumor plasma membrane chloroform-methanol (2:1, v/v) extract. Phosphorylation of protein was not affected by cAMP or cGMP. However, calcium ion (in the presence or absence of calmodulin) significantly modifies the 32P labelling of a series of proteins in normal tissue but has little effect with the neoplastic preparations. Some plasma membrane proteins were capable of nucleotide binding, instead or in addition to being phosphorylated. Finally, the presence of membrane-bound phosphoprotein phosphatase(s) was also demonstrated in all the preparations examined by means of chase experiments with nonlabelled ATP or GTP, and (or) by the use of the phosphoprotein phosphatase inhibitor, orthovanadate.
Church
J G
JG
Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, Canada.
Ghosh
S
S
Roufogalis
B D
BD
Villalobo
A
A
D000255:Adenosine Triphosphate / Q000378:metabolism; D000375:Aging; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D006160:Guanosine Triphosphate / Q000378:metabolism; D008099:Liver / Q000201:enzymology / Q000254:growth & development; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D008115:Liver Regeneration; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D008563:Membrane Lipids; D008565:Membrane Proteins; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D011494:Protein Kinases