Interfacial binding dynamics of bee venom phospholipase A2 investigated by dynamic light scattering and quartz crystal microbalance.
Bee venom phospholipase A(2) (bvPLA(2)) is part of the secretory phospholipase A(2) (sPLA(2)) family whose members are active in biological processes such as signal transduction and lipid metabolism. While controlling sPLA(2) activity is of pharmaceutical interest, the relationship between their mechanistic actions and physiological functions is not well understood. Therefore, we investigated the interfacial binding process of bvPLA(2) to characterize its biophysical properties and gain insight into how membrane binding affects interfacial activation. Attention was focused on the role of membrane electrostatics in the binding process. Although dynamic light scattering experiments indicated that bvPLA(2) does not lyse lipid vesicles, a novel, nonhydrolytic activity was discovered. We employed a supported lipid bilayer platform on the quartz crystal microbalance with dissipation sensor to characterize this bilayer-disrupting behavior and determined that membrane electrostatics influence this activity. The data suggest that (1) adsorption of bvPLA(2) to model membranes is not primarily driven by electrostatic interactions; (2) lipid desorption can follow bvPLA(2) adsorption, resulting in nonhydrolytic bilayer-disruption; and (3) this desorption is driven by electrostatic interactions. Taken together, these findings provide evidence that interfacial binding of bvPLA(2) is a dynamic process, shedding light on how membrane electrostatics can modulate interfacial activation.
Jackman
Joshua A
JA
Department of Chemistry, University of Florida, Gainesville, Florida 32611, USA.
Cho
Nam-Joon
NJ
Duran
Randolph S
RS
Frank
Curtis W
CW
D001514:Bee Venoms / Q000201:enzymology*; D008027:Light; D008051:Lipid Bilayers; D054467:Phospholipases A2 / Q000378:metabolism*; D011791:Quartz; D012542:Scattering, Radiation; D055672:Static Electricity
D001514:Bee Venoms; D008051:Lipid Bilayers; D011791:Quartz; D054467:Phospholipases A2