Protein kinase C isoforms in normal and chronic myeloid leukemic neutrophils. Distinct signal for PKC alpha by immunodetection on PVDF membrane, decreased expression of PKC alpha and increased expression of PKC delta in leukemic neutrophils.
Neutrophils from patients with Chronic Myeloid Leukemia (CML) exhibit defects in several functions. They also show altered phosphorylation-dephosphorylation patterns of several proteins on stimulation with phorbol myristate acetate--a direct activator of protein kinase C (PKC). Since PKC mediates several functions of the neutrophil, in this study we investigate the PKC isoform profile and subcellular distribution in normal and CML neutrophils in an attempt to elucidate their role in CML signalling. Our results show the presence of PKC alpha, betaI, betaII and delta in both the cell types. A distinct and clear signal was obtained for PKC alpha, the isoform reported to be absent or present in very low amounts in normal neutrophils. In addition, PKC alpha was present in significantly lower levels in CML neutrophils while the PKC delta isoform was found in significantly higher amounts in the CML cytosol as compared to that in normal cells. PKC alpha, betaI, betaII and delta isoforms could not be detected in the nucleus of unstimulated normal and CML neutrophils. The altered levels of PKC alpha and delta may be one of the causes for the defects in function exhibited by the leukemic cells.
Molecular Biology Unit, Cancer Research Institute, Parel, Mumbai, India.
N
N
Balasubramanian
S H
SH
Advani
S M
SM
Zingde
D000906:Antibodies / Q000276:immunology; D001672:Biocompatible Materials / Q000737:chemistry; D015153:Blotting, Western; D002458:Cell Fractionation; D003101:Collodion; D003600:Cytosol / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry; D007527:Isoenzymes / Q000032:analysis / Q000096:biosynthesis / Q000097:blood* / Q000737:chemistry* / Q000276:immunology; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood / Q000473:pathology*; D008163:Luminescent Measurements; D008567:Membranes, Artificial; D009504:Neutrophils / Q000737:chemistry* / Q000378:metabolism; D011145:Polyvinyls; D011493:Protein Kinase C / Q000032:analysis / Q000096:biosynthesis / Q000097:blood* / Q000737:chemistry* / Q000276:immunology; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta; D011506:Proteins / Q000032:analysis; D012967:Sodium Dodecyl Sulfate; D013010:Sonication
D000906:Antibodies; D001672:Biocompatible Materials; D007527:Isoenzymes; D008567:Membranes, Artificial; D011145:Polyvinyls; D011506:Proteins; C077776:polyvinyl fluoride; D012967:Sodium Dodecyl Sulfate; D003101:Collodion; C496628:PRKCA protein, human; C497258:PRKCD protein, human; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta