CHARACTERIZATION OF PROTEIN-KINASE-C ISOTYPE EXPRESSION IN ADULT-RAT HEART - PROTEIN-KINASE C-EPSILON IS A MAJOR ISOTYPE PRESENT, AND IT IS ACTIVATED BY PHORBOL ESTERS, EPINEPHRINE, AND ENDOTHELIN
CHARACTERIZATION OF PROTEIN-KINASE-C ISOTYPE EXPRESSION IN ADULT-RAT HEART - PROTEIN-KINASE C-EPSILON IS A MAJOR ISOTYPE PRESENT, AND IT IS ACTIVATED BY PHORBOL ESTERS, EPINEPHRINE, AND ENDOTHELIN
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DOI：
10.1161/01.res.72.4.757
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发表时间：
1993-04-01
影响因子：
20.1
通讯作者：
SUGDEN, PH
中科院分区：
文献类型：
--
作者：
BOGOYEVITCH, MA;PARKER, PJ;SUGDEN, PH
The pattern of protein kinase C (PKC) isotype expression in whole extracts of dispersed, freshly isolated adult rat ventricular myocytes and adult rat heart ventricle was examined by immunoblot analysis using antisera specific for PKC-alpha, -beta1, -gamma, -delta, -epsilon, -zeta, or -eta isotypes. This analysis revealed significant levels of expression of the Ca2+-independent isotype PKC-epsilon, which was detected as a band of 97-kd molecular mass. PKC-zeta was detected principally as a 66-kd band that probably represented a proteolytic product of the holoenzyme. PKC-eta was detected only in whole ventricle as a doublet at 75 and 81 kd and was therefore probably present in nonmyocytic cells. PKC-alpha, -beta1, -gamma, and -delta could not be detected. Because of our inability to detect PKC-alpha, -beta1, -gamma, and -delta in whole extracts, PKC isotypes were partially purified from whole heart by DEAE Sepharose chromatography. PKC-alpha, -beta1, -gamma, and -delta could still not be detected in the appropriate fractions. All PKC isotypes were detectable in appropriate positive control extracts (brain or certain cultured cell lines). In unstimulated isolated cardiomyocytes, the majority (80-95%) of the PKC-epsilon immunoreactivity was present in the soluble fraction of the extract. On exposure of the cardiomyocytes to 1 muM phorbol 12-myristate 13-acetate (PMA), PKC-epsilon undergoes a rapid (
