title: Direct observations of the cleavage reaction of an L-DPPC monolayer catalyzed by phospholipase A2 and inhibited by an indole inhibitor at the air/water interface.
abstract: The enzymatic hydrolysis of an L-dipalmitoylphosphatidylcholine (L-DPPC) monolayer at the air/water interface, catalyzed by phospholipase A(2) (PLA(2)), serves as a model for biospecific interfacial reactions. The cleavage of L-DPPC was investigated by Brewster angle microscopy. Different types of domain defects were observed to form in the coexisting liquid expanded and liquid condensed phases during the hydrolysis reaction. The adsorption of the enzyme was quantitatively recorded as the increase of the surface pressure over a fixed molecular area with time. In the case of L-DPPC, the surface pressure first increases and then starts to decrease, which indicates that a soluble product (lysolipid) is produced during the catalytic cleavage reaction. The increase and decrease of the surface pressure, which corresponds to the change of shape and number density of domains, indicated the occurrence of the following processes: adsorption of PLA(2), cleavage reaction, and rearrangement of substrate and product molecules at the interface. Addition of a PLA(2) inhibitor to the lipid monolayer leads to a fast surface pressure increase after enzyme injection. The surface pressure reaches a maximum value and then does not change for a long time. During this period, no change in the domain shape and number density was observed, which indicates that the enzyme is inhibited for a certain period of time. The experimental results provide the possibility of a direct way to prove inhibitor activity.
authors.affiliation: The Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences Zhong Guan Cun, Beijing 100080, PR China. jbli@infoc3.icas.ac.cn
authors.forename: Xiuhong
authors.initials: X
authors.lastname: Zhai
authors.forename: Junbai
authors.initials: J
authors.lastname: Li
authors.forename: Gerald
authors.initials: G
authors.lastname: Brezesinski
authors.forename: Qiang
authors.initials: Q
authors.lastname: He
authors.forename: Helmuth
authors.initials: H
authors.lastname: Möhwald
authors.forename: Luhua
authors.initials: L
authors.lastname: Lai
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Liang
authors.initials: L
authors.lastname: Liu
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry*; D000388:Air; D002384:Catalysis; D006868:Hydrolysis; D007211:Indoles / Q000737:chemistry*; D008853:Microscopy; D008954:Models, Biological; D008958:Models, Molecular; D015394:Molecular Structure; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000737:chemistry*; D054467:Phospholipases A2; D011312:Pressure; D013499:Surface Properties; D014867:Water; D014961:X-Ray Diffraction
chemical_list: D007211:Indoles; D014867:Water; D015060:1,2-Dipalmitoylphosphatidylcholine; D010741:Phospholipases A; D054467:Phospholipases A2