Phospholipase A2: A Paradigm of Allosteric Regulation by Membranes

Defining the molecular details and consequences of the association of water-soluble proteins with membranes is fundamental to understanding protein–lipid interactions and membrane functioning. Phospholipase A2 (PLA2) enzymes, which catalyze the hydrolysis of phospholipid substrates that comprise the membrane bilayer, provide the ideal system for studying protein–lipid interactions. Our research focuses on understanding the catalytic cycle of various recombinant human intracellular PLA2 enzymes including the GIVA cPLA2 and GVIA iPLA2. cPLA2 is the main arachidonic acid provider for the eicosanoid pathway, while iPLA2 is involved in membrane phospholipid remodeling.

Hydrogen/deuterium exchange mass spectrometry was used to experimentally identify distinct PLA2 peptide regions that interact with phospholipid vesicles. Molecular dynamics simulations, guided by the experimental data, showed that the active sites of these enzymes are allosterically regulated by membranes. Membrane phospholipids bind to allosteric sites located on the interfacial surface of PLA2s shifting their conformation from the “closed” to the “open” state. This process facilitates extraction and binding of a phospholipid molecule in the active site where the hydrolysis occurs. This constitutes the first detailed study describing the binding and interaction mechanism of PLA2s with the membrane bilayer as well as how they bind a single phospholipid molecule in the catalytic site. These enzymes are implicated in chronic inflammatory diseases and understanding their association with membranes, mechanism of action and interactions at the molecular level, will allow us to identify potent and selective inhibitors that can be further developed as anti-inflammatory agents.

Video Simulations

Mov.2 iPLA2SMDHighRes.mpg

Extraction of PAPC substrate molecule from the membrane into the GVIA iPLA2 pocket using a steered MD simulation. Copyright © 2017 National Academy of Sciences

Mov.6 iPLA2PAPCPOPCHighRes.mpg

MD simulation of the GVIA iPLA2–PAPC complex showing the POPC molecules in the membrane patch near the enzyme binding pocket. Copyright © 2017 National Academy of Sciences

Mov.9 iPLA2PAPCPocVol.mpg

Volume of the binding pocket of GVIA iPLA2 during the MD simulation. Copyright © 2017 National Academy of Sciences

Awards

The picture shows Dr. Mouchlis receiving the award from Prof. Carlos Simmerling in the presence of his mentor, Prof. Edward A. Dennis.


Dr. Varnavas D. Mouchlis presented his work at the 254th ACS National Meeting in San Francisco where he won the Wiley Computers in Chemistry Outstanding Postdoc Award.

http://www.acscomp.org/awards/the-comp-acs-outstanding-postdoc-award

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