title: Ligand-induced conformational changes in cytosolic protein kinase C.
abstract: The changes in intrinsic spectral properties of protein kinase C were monitored upon association with its divalent cation and lipid activators in a model membrane system. The enzyme demonstrated changes in both its intrinsic fluorescence and far ultraviolet circular dichroism spectra upon association with lipid vesicles in the absence of calcium. The acidic phospholipid, phosphatidylserine, significantly quenched the intrinsic tryptophan fluorescence and was also the most potent lipid support for the phosphorylating activity of the enzyme. The enzyme was fully activated by a number of Ca2(+)-lipid combinations which correlated with maximal fluorescence quenching (40-50%) of available tryptophan residues in hydrophobic domains. The circular dichroism structure of the associated active-protein Ca2(+)-lipid complexes suggested different active enzyme secondary structures. However, the Ca2(+)-dependent changes in fluorescence and circular dichroism spectra were observed only after the enzyme associated with the lipid vesicles. These data suggest that protein kinase C has the properties of a complex multidomain protein and provides an additional perspective into the mechanism of protein kinase C activation.
authors.affiliation: Department of Membrane Research, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: D S
authors.initials: DS
authors.lastname: Lester
authors.forename: V
authors.initials: V
authors.lastname: Brumfeld
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology; D002118:Calcium / Q000378:metabolism; D002942:Circular Dichroism; D003600:Cytosol / Q000201:enzymology*; D008024:Ligands; D050356:Lipid Metabolism; D008274:Magnesium / Q000378:metabolism; D008297:Male; D011487:Protein Conformation; D011493:Protein Kinase C / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence
chemical_list: D008024:Ligands; D011493:Protein Kinase C; D008274:Magnesium; D002118:Calcium