Signal transduction mechanism in human neutrophil: comparative study between the zeta and beta-protein kinase isotypes.
Role of protein kinase C (PKC) isotypes in the regulation of neutrophil function are not clearly known. In the present study we purified the beta-PKC and zeta-PKC isotypes from human neutrophil. Both the isotypes are immunoreactive only to their respective antibodies. Zeta-PKC was further confirmed by RT-PCR using specific primer. Co-factor requirements for both the kinases were found to be different when DG and ceramide were used as second messenger. Selective substrate specificities were determined for both beta and zeta-PKC using isotype specific pseudosubstrates viz., [Ser25]PKC[19-31] and [Ser119]PKC[113-130] respectively. Endogenous protein phosphorylation by purified beta-PKC and zeta-PKC showed their functional differences in neutrophil. Beta-PKC phosphorylated 13, 15, 19, 33, 36, 47, 80 and 92 kDa proteins and zeta-PKC phosphorylated 19, 22, 42, 47, 75 and 87 kDa proteins, only exception was the phosphorylation of 47 kDa protein which had been phosphorylated by both the kinases. Differences in phosphorylation between beta-PKC and zeta-PKC clearly indicate the selective role for these PKC isotypes in the activation sequences of neutrophil.
Das
S
S
Department of Microbiology, Bose Institute, Calcutta, India.
Bhattacharyya
S
S
Ghosh
S
S
Majumdar
S
S
D001483:Base Sequence; D017931:DNA Primers; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D064546:Protein Kinase C beta; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013379:Substrate Specificity; D000099221:Protein Kinase C zeta
D017931:DNA Primers; D007527:Isoenzymes; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D000099221:Protein Kinase C zeta