Characterization of 130 kDa protein from rat cerebellum synaptosomal membranes phosphorylated by PKC.
1. The effect of endogenous PMA-stimulated phosphorylation of the protein in the molecular weight range of 130 kDa in rat cerebellum synaptosomal membranes was examined. 2. The 50% inhibition of the phosphorylation of 130 kDa protein by 5 microM polymyxin B was observed after 6 min of preincubation. 3. The sensitivity of 130 kDa protein for phosphorylation in the presence of exogenous protein kinase C suggests, that this protein could serve as a physiological substrate of protein kinase C. 4. Partial characterization of 130 kDa protein was performed. Upon incubation with [gamma-32P]ATP the 130 kDa protein formed Ca(2+)-dependent, hydroxylamine-sensitive phosphointermediate, which was inhibited by 50 microM vanadate, but not 0.5 mM vanadyl. 5. One-dimensional peptide mapping by proteolysis of 130 kDa protein with V8 protease was obtained.
Zylińska
L
L
II Department of Biochemistry, Institute of Physiology and Biochemistry, School of Medicine, Lódź, Poland.
Lachowicz
L
L
D000818:Animals; D002531:Cerebellum / Q000737:chemistry*; D007425:Intracellular Membranes / Q000737:chemistry*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D010449:Peptide Mapping; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012697:Serine Endopeptidases; D013574:Synaptosomes / Q000737:chemistry*; D014638:Vanadates / Q000494:pharmacology
D009419:Nerve Tissue Proteins; D014638:Vanadates; D011493:Protein Kinase C; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase