Structure of the C2 domain from novel protein kinase Cϵ. A membrane binding model for Ca2+-independent C2 domains

2001 ◽  
Vol 311 (4) ◽  
pp. 837-849 ◽  
Author(s):  
Wendy F Ochoa ◽  
Josefa Garcia-Garcia ◽  
Ignacio Fita ◽  
Senena Corbalan-Garcia ◽  
Nuria Verdaguer ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Membrane Binding ◽  
C2 Domain ◽  
Binding Model ◽  
C2 Domains ◽  
Novel Protein

scholarly journals Physiological Role for Phosphatidic Acid in the Translocation of the Novel Protein Kinase C Apl II in Aplysia Neurons

2008 ◽  
Vol 28 (15) ◽  
pp. 4719-4733 ◽  
Author(s):  
Carole A. Farah ◽  
Ikue Nagakura ◽  
Daniel Weatherill ◽  
Xiaotang Fan ◽  
Wayne S. Sossin
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phosphatidic Acid ◽  
Physiological Role ◽  
C2 Domain ◽  
The Novel ◽  
Physiological Conditions ◽  
Kinase C ◽  
Neuronal Membranes ◽  
Novel Protein

ABSTRACT In Aplysia californica, the serotonin-mediated translocation of protein kinase C (PKC) Apl II to neuronal membranes is important for synaptic plasticity. The orthologue of PKC Apl II, PKCε, has been reported to require phosphatidic acid (PA) in conjunction with diacylglycerol (DAG) for translocation. We find that PKC Apl II can be synergistically translocated to membranes by the combination of DAG and PA. We identify a mutation in the C1b domain (arginine 273 to histidine; PKC Apl II-R273H) that removes the effects of exogenous PA. In Aplysia neurons, the inhibition of endogenous PA production by 1-butanol inhibited the physiological translocation of PKC Apl II by serotonin in the cell body and at the synapse but not the translocation of PKC Apl II-R273H. The translocation of PKC Apl II-R273H in the absence of PA was explained by two additional effects of this mutation: (i) the mutation removed C2 domain-mediated inhibition, and (ii) the mutation decreased the concentration of DAG required for PKC Apl II translocation. We present a model in which, under physiological conditions, PA is important to activate the novel PKC Apl II both by synergizing with DAG and removing C2 domain-mediated inhibition.

scholarly journals Specific Translocation of Protein Kinase Cα to the Plasma Membrane Requires Both Ca2+ and PIP2 Recognition by Its C2 Domain

2006 ◽  
Vol 17 (1) ◽  
pp. 56-66 ◽  
Author(s):  
John H. Evans ◽  
Diana Murray ◽  
Christina C. Leslie ◽  
Joseph J. Falke
Keyword(s):  
Plasma Membrane ◽  
Protein Kinase ◽  
Membrane Binding ◽  
Membrane Lipid ◽  
C2 Domain ◽  
Plasma Membrane Lipid ◽  
Protein Kinase Cα ◽  
Golgi Network

The C2 domain of protein kinase Cα (PKCα) controls the translocation of this kinase from the cytoplasm to the plasma membrane during cytoplasmic Ca2+ signals. The present study uses intracellular coimaging of fluorescent fusion proteins and an in vitro FRET membrane-binding assay to further investigate the nature of this translocation. We find that Ca2+-activated PKCα and its isolated C2 domain localize exclusively to the plasma membrane in vivo and that a plasma membrane lipid, phosphatidylinositol-4,5-bisphosphate (PIP2), dramatically enhances the Ca2+-triggered binding of the C2 domain to membranes in vitro. Similarly, a hybrid construct substituting the PKCα Ca2+-binding loops (CBLs) and PIP2 binding site (β-strands 3–4) into a different C2 domain exhibits native Ca2+-triggered targeting to plasma membrane and recognizes PIP2. Conversely, a hybrid containing the CBLs but lacking the PIP2 site translocates primarily to trans-Golgi network (TGN) and fails to recognize PIP2. Similarly, PKCα C2 domains possessing mutations in the PIP2 site target primarily to TGN and fail to recognize PIP2. Overall, these findings demonstrate that the CBLs are essential for Ca2+-triggered membrane binding but are not sufficient for specific plasma membrane targeting. Instead, targeting specificity is provided by basic residues on β-strands 3–4, which bind to plasma membrane PIP2.

The C2 domains of classical/conventional PKCs are specific PtdIns(4,5)P2-sensing domains

2007 ◽  
Vol 35 (5) ◽  
pp. 1046-1048 ◽  
Author(s):  
S. Corbalán-García ◽  
M. Guerrero-Valero ◽  
C. Marín-Vicente ◽  
J.C. Gómez-Fernández
Keyword(s):  
Plasma Membrane ◽  
Protein Kinase ◽  
Living Cells ◽  
Model Membranes ◽  
Membrane Localization ◽  
Dependent Manner ◽  
C2 Domain ◽  
C2 Domains ◽  
Local Densities

The C2 domains of cPKCs [classical/conventional PKCs (protein kinase Cs)] bind to membranes in a Ca2+-dependent manner and thereby act as cellular Ca2+ effectors. Recent findings have demonstrated that the C2 domain of cPKCs interacts specifically with PtdIns(4,5)P2 through its polybasic cluster located in the β3–β4-strands, this interaction being critical for the membrane localization of these enzymes in living cells. In addition, these C2 domains exhibit higher affinity to bind PtdIns(4,5)P2 than any other polyphosphate phosphatidylinositols. It has also been shown that the presence of PtdIns(4,5)P2 in model membranes decreases the Ca2+ concentration required for classical C2 domains to bind them. Overall, the studies reviewed here suggest a new mechanism of membrane docking by the C2 domains of cPKCs in which the local densities of phosphatidylserine and PtdIns(4,5)P2 on the inner leaflet of the plasma membrane are sufficient to drive Ca2+-activated membrane docking during a physiological Ca2+ signal.

Roles of calcium ions in the membrane binding of C2 domains

2001 ◽  
Vol 359 (3) ◽  
pp. 679-685 ◽  
Author(s):  
Robert V. STAHELIN ◽  
Wonhwa CHO
Keyword(s):  
Conformational Changes ◽  
Membrane Trafficking ◽  
Electrostatic Interactions ◽  
Membrane Binding ◽  
The Other ◽  
Lipid Monolayer ◽  
Dependent Manner ◽  
C2 Domain ◽  
Membrane Penetration ◽  
C2 Domains

The C2 domain is a membrane-targeting domain found in many cellular proteins involved in signal transduction or membrane trafficking. The majority of C2 domains co-ordinate multiple Ca2+ ions and bind the membrane in a Ca2+-dependent manner. To understand the mechanisms by which Ca2+ mediates the membrane binding of C2 domains, we measured the membrane binding of the C2 domains of group IV cytosolic phospholipase A2 (cPLA2) and protein kinase C-α (PKC-α) by surface plasmon resonance and lipid monolayer analyses. Ca2+ ions mainly slow the membrane dissociation of cPLA2-C2, while modulating both membrane association and dissociation rates for PKC-α-C2. Further studies with selected mutants showed that for cPLA2 a Ca2+ ion bound to the C2 domain of cPLA2 induces the intra-domain conformational change that leads to the membrane penetration of the C2 domain whereas the other Ca2+ is not directly involved in membrane binding. For PKC-α, a Ca2+ ion induces the inter-domain conformational changes of the protein and the membrane penetration of non-C2 residues. The other Ca2+ ion of PKC-α-C2 is involved in more complex interactions with the membrane, including both non-specific and specific electrostatic interactions. Together, these studies of isolated C2 domains and their parent proteins allow for the determination of the distinct and specific roles of each Ca2+ ion bound to different C2 domains.

scholarly journals Autophosphorylation of the C2 domain inhibits translocation of the novel protein kinase C (nPKC) Apl II

2012 ◽  
Vol 123 (3) ◽  
pp. 360-372 ◽  
Author(s):  
Carole A. Farah ◽  
Amanda A. Lindeman ◽  
Vincent Siu ◽  
Micaela Das Gupta ◽  
Wayne S. Sossin
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
C2 Domain ◽  
The Novel ◽  
Kinase C ◽  
Novel Protein

Membrane Binding Kinetics of Protein Kinase C βII Mediated by the C2 Domain†

Biochemistry ◽  
2001 ◽  
Vol 40 (44) ◽  
pp. 13216-13229 ◽  
Author(s):  
Eric A. Nalefski ◽  
Alexandra C. Newton
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Membrane Binding ◽  
Binding Kinetics ◽  
C2 Domain ◽  
Kinase C ◽  
Kinetics Of

scholarly journals Structural characterization of the C2 domain of novel protein kinase Cε

2001 ◽  
Vol 268 (4) ◽  
pp. 1107-1117 ◽  
Author(s):  
Josefa García-García ◽  
Juan C. Gómez-Fernández ◽  
Senena Corbalán-García
Keyword(s):  
Protein Kinase ◽  
Structural Characterization ◽  
C2 Domain ◽  
Novel Protein
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