Mouse protein kinase C-delta, the major isoform expressed in mouse hemopoietic cells: sequence of the cDNA, expression patterns, and characterization of the protein.
A complementary DNA (cDNA) of 2559 bp which encode all 674 amino acids of mouse protein kinase C-delta (PKC-delta) has been isolated from a cDNA library prepared from ABPL-2, a mouse myeloid tumor. The library was screened with a partial PKC-delta cDNA clone that had been created by polymerase chain reaction (PCR) amplification of ABPL-2 RNA using primers that are conserved among all rat PKC isozymes. This approach proved to be a distinct improvement over screening with synthetic oligonucleotides. Similar sets of cDNAs prepared from other hemopoietic cell lines were screened with this PKC-delta cDNA and with probes for the other PKC isoforms. These experiments revealed that the major isoform of PKC expressed in hemopoietic cells is PKC-delta. PKC-delta protein was purified from ABPL-3, a mouse myeloid tumor which expressed principally the delta isoform of PKC. The protein eluted from a hydroxylapatite column in the same position as PKC-beta and -epsilon would elute, if present. The kinase activity of purified PKC-delta showed strict dependence on the presence of phospholipids, but showed no activation by Ca2+.
Mischak
H
H
Laboratory of Genetics, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Bodenteich
A
A
Kolch
W
W
Goodnight
J
J
Hofer
F
F
Mushinski
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D002852:Chromatography, Ion Exchange; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D015723:Gene Library; D007527:Isoenzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007942:Leukemia, Experimental; D007951:Leukemia, Myeloid; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011493:Protein Kinase C / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D004273:DNA, Neoplasm; D007527:Isoenzymes; D011994:Recombinant Proteins; D011493:Protein Kinase C