Redistribution of a 120 kDa phosphoprotein in the parietal cell associated with stimulation.
When rabbit isolated gastric glands were stimulated via the cyclic AMP pathway, a phosphorylated protein band of about 120 kDa (pp120) was markedly increased in the apical membrane-rich fraction, concomitant with an increase in the amount of H,K-ATPase and the phosphorylation of the cytoskeletal protein ezrin in the same fraction. The cytosolic fraction, but not other membrane fractions, also contained a protein with common features to the membrane-bound pp120, i.e., comigration in two-dimensional gels with a pI of approximately 4.5, anomalous mobility in SDS-PAGE, reactivity to antibodies, and phosphorylation, indicating that these two proteins were identical. The possibility that pp120 is vinculin was completely excluded. Using antibody against pp120, this protein was found to be almost exclusively in the gastric parietal cell. Biochemical and immunohistochemical analyses suggest that pp120 exists mainly in the cytosol, and that a small part of the protein binds to the apical membrane when the parietal cell is stimulated via the cyclic AMP pathway. In the presence of histone, purified pp120 produced phosphorylation on pp120 as well as histone. The inhibitor profile of this kinase activity is not consistent with any known kinase. We conclude that pp120 is closely associated with a new type of kinase, and translocates from cytosol to the apical membrane when the parietal cell is stimulated.
Urushidani
T
T
Department of Cell Biology, Institute of Basic Medical Sciences, University of Tsukuba.
Chow
D
D
Forte
J G
JG
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology*; D000818:Animals; D001692:Biological Transport; D003994:Bucladesine / Q000494:pharmacology*; D002462:Cell Membrane / Q000201:enzymology; D016764:Cell Polarity; D000242:Cyclic AMP / Q000502:physiology*; D017506:H(+)-K(+)-Exchanging ATPase / Q000378:metabolism; D006632:Histamine / Q000494:pharmacology*; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D010295:Parietal Cells, Gastric / Q000187:drug effects / Q000378:metabolism*; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D015290:Second Messenger Systems / Q000187:drug effects*; D016596:Vinculin / Q000302:isolation & purification / Q000378:metabolism
D010750:Phosphoproteins; D016596:Vinculin; D003994:Bucladesine; D006632:Histamine; D000242:Cyclic AMP; D011494:Protein Kinases; D017506:H(+)-K(+)-Exchanging ATPase; D015056:1-Methyl-3-isobutylxanthine