title: Phospholipase A2 hydrolysis of supported phospholipid bilayers: a neutron reflectivity and ellipsometry study.
abstract: We have investigated the phospholipase A(2) catalyzed hydrolysis of supported phospholipid bilayers using neutron reflection and ellipsometry. At the hydrophilic silica-water interface, hydrolysis of phosphatidylcholine bilayers by phospholipase A(2) from Naja mossambica mossambica venom is accompanied by destruction of the bilayer at an initial rate, which is comparable for DOPC and DPPC but is doubled for POPC. The extent of bilayer destruction at 25 degrees C decreases from DOPC to POPC and is dramatically reduced for DPPC. Neutron reflectivity measurements indicate that the enzyme penetrates into the bilayers in increasing order for DOPC, POPC, and DPPC, while the amount of enzyme adsorbed at the interface is smallest for DPPC and exhibits a maximum for POPC. Penetration into the hydrophobic chain region in the bilayer is further supported by the fact that the enzyme adsorbs strongly and irreversibly to a hydrophobic monolayer of octadecyltrichlorosilane. These results are rationalized in terms of the properties of the reaction products and the effect of their accumulation in the membrane on the kinetics of enzyme catalysis.
authors.affiliation: Oxford University, Physical and Theoretical Chemistry Laboratory, South Parks Road, Oxford OX1 3QZ, United Kingdom. hanna.vacklin@chem.ox.ac.uk
authors.forename: Hanna P
authors.initials: HP
authors.lastname: Vacklin
authors.forename: Fredrik
authors.initials: F
authors.lastname: Tiberg
authors.forename: Giovanna
authors.initials: G
authors.lastname: Fragneto
authors.forename: R K
authors.initials: RK
authors.lastname: Thomas
mesh_terms: D000818:Animals; D004546:Elapid Venoms; D017815:Elapidae; D006868:Hydrolysis; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism*; D008958:Models, Molecular; D009502:Neutrons; D010741:Phospholipases A / Q000737:chemistry* / Q000378:metabolism*; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism*; D011487:Protein Conformation; D013499:Surface Properties
chemical_list: D004546:Elapid Venoms; D008051:Lipid Bilayers; D010743:Phospholipids; D010741:Phospholipases A; D054467:Phospholipases A2