Protein kinase C mu is located at the Golgi compartment.
Protein kinase C mu (PKC mu) displays unusual structural features like a pleckstrin homology domain and an amino-terminal hydrophobic region with a putative leader peptide and transmembrane sequence. As a discrete location often is a direct clue to the potential biological function of a kinase, antibodies directed against unique amino- and carboxy-terminal domains of PKC mu were used to localize the protein within intracellular compartments in immunofluorescence and subcellular fractionation studies. Confocal laser scanning microscopy showed colocalization of PKC mu with the resident Golgi marker protein beta 1,4 galactosyltransferase in PKC mu transfectants and in the human hepatocellular carcinoma cell line HepG2, expressing endogenous PKC mu. Long-term treatment of cells with brefeldin A, which disintegrates the Golgi apparatus, disrupted PKC mu-specific staining. Cosegregation of PKC mu with beta 1,4 galactosyltransferase, but not with the endosomal marker rab5, upon density gradient fractionation and Western blot analysis of HepG2 cell extracts, provides independent evidence for a Golgi localization of PKC mu. Moreover, cellular sulfate uptake and Golgi-specific glycosaminoglycan sulfation was enhanced in PKC mu transfectants. Together, these data suggest that PKC mu is a resident protein kinase of the core Golgi compartment and is involved in basal transport processes.
Prestle
J
J
Institute of Cell Biology and Immunology, University of Stuttgart, Germany.
Pfizenmaier
K
K
Brenner
J
J
Johannes
F J
FJ
D016475:3T3 Cells / Q000201:enzymology; D000818:Animals; D000918:Antibody Specificity; D015152:Blotting, Northern; D016466:CHO Cells / Q000201:enzymology; D006224:Cricetinae; D019084:Fluorescent Antibody Technique, Indirect; D005700:Galactosyltransferases; D006025:Glycosaminoglycans / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D011493:Protein Kinase C / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D013347:Subcellular Fractions / Q000201:enzymology; D013431:Sulfates / Q000378:metabolism; D014162:Transfection
D006025:Glycosaminoglycans; D007527:Isoenzymes; D012333:RNA, Messenger; D013431:Sulfates; D005700:Galactosyltransferases; D011493:Protein Kinase C