title: Increased membrane affinity of the C1 domain of protein kinase Cdelta compensates for the lack of involvement of its C2 domain in membrane recruitment.
abstract: Protein kinase C (PKC) family members are allosterically activated following membrane recruitment by specific membrane-targeting modules. Conventional PKC isozymes are recruited to membranes by two such modules: a C1 domain, which binds diacylglycerol (DAG), and a C2 domain, which is a Ca2+-triggered phospholipid-binding module. In contrast, novel PKC isozymes respond only to DAG, despite the presence of a C2 domain. Here, we address the molecular mechanism of membrane recruitment of the novel isozyme PKCdelta. We show that PKCdelta and a conventional isozyme, PKCbetaII, bind membranes with comparable affinities. However, dissection of the contribution of individual domains to this binding revealed that, although the C2 domain is a major determinant in driving the interaction of PKCbetaII with membranes, the C2 domain of PKCdelta does not bind membranes. Instead, the C1B domain is the determinant that drives the interaction of PKCdelta with membranes. The C2 domain also does not play any detectable role in the activity or subcellular location of PKCdelta in cells; in vivo imaging studies revealed that deletion of the C2 domain does not affect the stimulus-dependent translocation or activity of PKCdelta. Thus, the increased affinity of the C1 domain of PKCdelta allows this isozyme to respond to DAG alone, whereas conventional PKC isozymes require the coordinated action of Ca2+ binding to the C2 domain and DAG binding to the C1 domain for activation.
authors.affiliation: Department of Pharmacology, University of California at San Diego, La Jolla, California 92093-0721, USA.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Giorgione
authors.forename: Jung-Hsin
authors.initials: JH
authors.lastname: Lin
authors.forename: J Andrew
authors.initials: JA
authors.lastname: McCammon
authors.forename: Alexandra C
authors.initials: AC
authors.lastname: Newton
mesh_terms: D001665:Binding Sites; D002462:Cell Membrane / Q000201:enzymology*; D017931:DNA Primers; D004075:Diglycerides / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D051745:Protein Kinase C-delta / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013816:Thermodynamics
chemical_list: D017931:DNA Primers; D004075:Diglycerides; D008081:Liposomes; D010743:Phospholipids; D011994:Recombinant Proteins; D051745:Protein Kinase C-delta; D013755:Tetradecanoylphorbol Acetate