Protein kinase C bound to the Golgi apparatus supports the formation of constitutive transport vesicles

被引：54

作者：

Westermann, P ^{[1
]}

Knoblich, M ^{[1
]}

Maier, O ^{[1
]}

Lindschau, C ^{[1
]}

Haller, H ^{[1
]}

机构：

[1] HUMBOLDT UNIV BERLIN,VIRCHOW KLINIKUM,FRANZ VOLHARD CLIN,D-13125 BERLIN,GERMANY

来源：

BIOCHEMICAL JOURNAL | 1996年 / 320卷

关键词：

D O I：

10.1042/bj3200651

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Constitutive secretion of heparan sulphate proteoglycans (HSPGs) was stimulated in human hepatoma HepG2 cells by phorbol 12-myristate 13-acetate (PMA) and inhibited by calphostin C, a specific inhibitor of protein kinase C (PKC). To delineate more closely the site of PKC action, the packaging in vitro of (SO4)-S-35-labelled HSPGs into transport vesicles was investigated. Formation of transport vesicles at the trans-Golgi network was stimulated by PMA and inhibited by calphostin C or Ro 318220 by using a post-nuclear supernatant. Treatment of either isolated Golgi-enriched membranes or cytosolic proteins with calphostin C provided evidence that membrane-bound PKC forms strongly supported vesicle formation, whereas cytosolic PKC forms showed a marginal effect. The PKC isoforms PKC-alpha and PKC-zeta were attached to highly purified Golgi membranes, as shown by Western blotting. Both isoforms were localized by confocal immunofluorescence microscopy in the Golgi area of HepG2 cells. Immunoelectron microscopy of ultrathin cryosections of HepG2 cells showed that PKC-zeta predominantly attaches to the trans-Golgi region, whereas PKC-a binds to the cis- and trans-Golgi area.

引用

页码：651 / 658

页数：8

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