A constant area monolayer method to assess optimal lipid packing for lipolysis tested with several secreted phospholipases A2

YUNES QUARTINO PJ.,; PORTELA M.,; LIMA A.,; DURAN R.,; LOMONTE B.,; FIDELIO G.D.

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2015 vol. 1848 p. 2216 - 2216

e present an analysis of lipid monolayer hydrolysis at a constant area to assess the optimal lateral surface pressure value (Πopt) and thus, the surface packing density of the lipid, at which the activity of a given lipolytic enzyme is maximal. This isochoricmethod consists of a measurement of the decrease down to zero of theΠopt of phospholipid substrate monolayer due to continuous hydrolysis using only one reaction compartment. Weperformed the comparison of both approaches using several commercially available and literature-evaluated sPLA2s. Also, we characterized for the first time the profile of hydrolysis of DLPC monolayers catalyzed by a sPLA2 from Streptomyces violaceoruber and isoenzymes purified from Bothrops diporus venom. One of these viper venomenzymes is a newisoenzyme, partially sequenced by amass spectrometry approach.We also included the basicmyotoxin sPLA2-III fromBothrops asper. Results obtained with the isochoricmethod and the standard isobaric one p