Membrane environment exerts an important influence on rac-mediated activation of phospholipase Cγ2.
We performed analyses of the molecular mechanisms involved in the regulation of phospholipase Cγ2 (PLCγ2). We identified several regions in the PLCγ-specific array, γSA, that contribute to autoinhibition in the basal state by occlusion of the catalytic domain. While the activation of PLCγ2 by Rac2 requires stable translocation to the membrane, the removal of the domains required for membrane translocation in the context of an enzyme with impaired autoinhibition generated constitutive, highly active PLC in cells. We further tested the possibility that the interaction of PLCγ2 with its activator protein Rac2 was sufficient for activation through the release of autoinhibition. However, we found that Rac2 binding in the absence of lipid surfaces was not able to activate PLCγ2. Together with other observations, these data suggest that an important consequence of Rac2 binding and translocation to the membrane is that membrane proximity, on its own or together with Rac2, has a role in the release of autoinhibition, resulting in interfacial activation.
Section of Cell and Molecular Biology, Chester Beatty Laboratories, The Institute of Cancer Research, Fulham Road, London SW3 6JB, United Kingdom.
Katy L
KL
Everett
Anja
A
Buehler
Tom D
TD
Bunney
Anca
A
Margineanu
Rhona W
RW
Baxendale
Petra
P
Vatter
Michael
M
Retlich
Claudia
C
Walliser
Hugh B
HB
Manning
Mark A A
MA
Neil
Christopher
C
Dunsby
Paul M W
PM
French
Peter
P
Gierschik
Matilda
M
Katan
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004789:Enzyme Activation; D006801:Humans; D051966:Phospholipase C gamma / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D020744:rac GTP-Binding Proteins / Q000378:metabolism*
D051966:Phospholipase C gamma; D020744:rac GTP-Binding Proteins
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