A comparison of the membrane binding properties of C1B domains of PKCgamma, PKCdelta, and PKCepsilon.
The C1 domains of classical and novel PKCs mediate their diacylglycerol-dependent translocation. Using fluorescence resonance energy transfer, we studied the contribution of different negatively charged phospholipids and diacylglycerols to membrane binding. Three different C1B domains of PKCs were studied (the classical gamma, and the novel delta and epsilon), together with different lipid mixtures containing three types of acidic phospholipids and three types of activating diacylglycerols. The results show that C1Bgamma and C1Bepsilon exhibit a higher affinity to bind to vesicles containing 1-palmitoyl-2-oleoyl-sn-phosphatidic acid, 1-palmitoyl-2-oleoyl-sn-phoshatidylserine, or 1-palmitoyl-2-oleoyl-sn-phosphatidylglycerol, with C1Bepsilon being the most relevant case because its affinity for POPA-containing vesicles increased by almost two orders of magnitude. When the effect of the diacylglycerol fatty acid composition on membrane binding was studied, the C1Bepsilon domain showed the highest binding affinity to membranes containing 1-stearoyl-oleoyl-sn-glycerol or 1,2-sn-dioleoylglycerol with POPA as the acidic phospholipid. Of the three diacylglycerols used in this study, 1,2-sn-dioleoylglycerol and 1-stearoyl-oleoyl-sn-glycerol showed the highest affinities for each isoenzyme, whereas 1,2-sn-dipalmitoylglycerol; showed the lowest affinity. DSC experiments showed this to be a consequence of the nonfluid conditions of 1,2-sn-dipalmitoylglycerol;-containing systems.
Departamento de Bioquímica y Biología Molecular A, Facultad de Veterinaria, Universidad de Murcia, E-30080-Murcia, Spain.
Sonia
S
Sánchez-Bautista
Senena
S
Corbalán-García
Angel
A
Pérez-Lara
Juan C
JC
Gómez-Fernández
D000241:Adenosine / Q000031:analogs & derivatives / Q000378:metabolism; D002152:Calorimetry, Differential Scanning; D002460:Cell Line; D004075:Diglycerides / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D020404:Glycerophospholipids / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D010712:Phosphatidic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D051745:Protein Kinase C-delta / Q000737:chemistry / Q000378:metabolism*; D051744:Protein Kinase C-epsilon / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013696:Temperature; D053835:Unilamellar Liposomes / Q000378:metabolism*
C103238:1-palmitoyl-2-oleoyl-glycero-3-phosphatidic acid; C509561:1-palmitoyl-2-oleoyl-phosphatidyl-3-adenosine; D004075:Diglycerides; D020404:Glycerophospholipids; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D010718:Phosphatidylserines; D053835:Unilamellar Liposomes; C050294:1-palmitoyl-2-oleoylglycero-3-phosphoserine; C084199:1-stearoyl-2-oleoyl-sn-glycerol; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; C087857:protein kinase C gamma; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D000241:Adenosine; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; C013965:diolein