PKC epsilon is associated with myosin IIA and actin in fibroblasts.
Proteins coimmunoprecipitating with protein kinase C (PKC) epsilon in fibroblasts were identified through matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI TOF m/s). This method identified myosin IIA in PKC epsilon immunoprecipitates, as well as known PKC epsilon binding proteins, actin, beta'Cop and cytokeratin. Myosin is not a substrate for PKC epsilon. Immunofluorescence analysis showed that PKC epsilon is colocalised with actin and myosin in actomyosin stress fibers in fibroblasts. Inhibitors of PKC and myosin ATPase activity, as well as microfilament-disrupting drugs, all inhibited spreading of fibroblasts after passage, suggesting a role for a PKC epsilon-actin-myosin complex in cell spreading.
England
Karen
K
Department of Biology, University of York, York YO10 5DD, UK. k.england@ucc.ie
Ashford
David
D
Kidd
Daniel
D
Rumsby
Martin
M
D016475:3T3 Cells; D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology* / Q000378:metabolism* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D007527:Isoenzymes / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051379:Mice; D024507:Nonmuscle Myosin Type IIA / Q000032:analysis / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051744:Protein Kinase C-epsilon; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D022502:Stress Fibers / Q000737:chemistry / Q000648:ultrastructure
D000199:Actins; D004791:Enzyme Inhibitors; D007527:Isoenzymes; C497256:Prkce protein, mouse; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D024507:Nonmuscle Myosin Type IIA