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Role of the Ca2+/phosphatidylserine binding region of the C2 domain in the translocation of protein kinase C alpha to the plasma membrane

Bolsover, SR; Gomez-Fernandez, JC; Corbalan-Garcia, S; (2003) Role of the Ca2+/phosphatidylserine binding region of the C2 domain in the translocation of protein kinase C alpha to the plasma membrane. J BIOL CHEM , 278 (12) 10282 - 10290. 10.1074/jbc.M212145200. Gold open access

Abstract

Signal transduction. via protein kinase C (PKC) is closely regulated by its subcellular localization. To map the molecular determinants mediating the C2 domain-dependent translocation of PKCalpha to the plasma membrane, full-length native protein and several point mutants in the Ca2+/phosphatidylserine-binding site were tagged with green fluorescent protein and transiently expressed in rat basophilic leukemia cells (RBL-2H3). Substitution of several aspartate residues by asparagine completely abolished Ca2+-dependent membrane targeting of PKCalpha. Strikingly, these mutations enabled the mutant proteins to translocate in a diacylglycerol-dependent manner, suggesting that neutralization of charges in the Ca2+ binding region enables the C1 domain to bind diacylglycerol. In addition, it was demonstrated that the protein residues involved in direct interactions with acidic phospholipids play differential and pivotal roles in the membrane targeting of the enzyme. These findings provide new information on how the C2 domain-dependent membrane targeting of PKCalpha occurs in the presence of physiological stimuli.

Type: Article
Title: Role of the Ca2+/phosphatidylserine binding region of the C2 domain in the translocation of protein kinase C alpha to the plasma membrane
Open access status: An open access publication
DOI: 10.1074/jbc.M212145200
Keywords: GREEN FLUORESCENT PROTEIN, SYNAPTOTAGMIN I, C(2)A DOMAIN, CALCIUM-IONS, CA2+ BINDING, BETA-II, ACTIVATION, DIACYLGLYCEROL, PHOSPHATIDYLSERINE, IDENTIFICATION
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology
URI: http://discovery.ucl.ac.uk/id/eprint/10247
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