Hydrolysis of mixed monomolecular films of tricaprylin/dilauroylphosphatidylcholine by lipase and phospholipase A2

We 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 isochoric method consists of a measurement of the decrease down to zero of the Πopt of phospholipid substrate monolayer due to continuous hydrolysis using only one reaction compartment. We performed 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 venom enzymes is a new isoenzyme, partially sequenced by a mass spectrometry approach. We also included the basic myotoxin sPLA2-III from Bothrops asper. Results obtained with the isochoric method and the standard isobaric one produced quite similar values of Πopt, validating the proposal. In addition, we propose a new classification parameter, a lipolytic ratio of hydrolysis at two lateral pressures, 20 mN·m(-1) and 10 mN·m(-1), termed here as LR20/10 index. This index differentiates quite well "high surface pressure" from "low surface pressure" sPLA2s and, by extension; it can be used as a functional criterion for the quality of a certain enzyme. Also, this index could be added to the grouping systematic criteria for the superfamily proposed for phospholipase A2.

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

Quartino

Portela

Lima

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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CNS myelin structural modification induced in vitro by phospholipases A2

Quartino

Pusterla

Josa

et al. 2016

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Myelin is the self-stacked membrane surrounding axons; it is also the target of several pathological and/or neurodegenerative processes like multiple sclerosis. These processes involve, among others, the hydrolytic attack by phospholipases. In this work we describe the changes in isolated myelin structure after treatment with several secreted PLA2 (sPLA2), by using small angle x-ray scattering (SAXS) measurements. It was observed that myelin treated with all the tested sPLA2s (from cobra and bee venoms and from pig pancreas) preserved the lamellar structure but displayed an enlarged separation between membranes in certain zones. Additionally, the peak due to membrane asymmetry was clearly enhanced. The coherence length was also lower than the non-treated myelin, indicating increased disorder. These SAXS results were complemented by Langmuir film experiments to follow myelin monolayer hydrolysis at the air/water interface by a decrease in electric surface potential at different surface pressures. All enzymes produced hydrolysis with no major qualitative difference between the isoforms tested.

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Action of Vipoxin and its separated components on monomolecular film of Dilauroylphosphatidylcholine at the air/water interface

Mircheva

Petrova

Ivanova

et al. 2019

Colloids and Surfaces A: Physicochemical and Engineering Aspect

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Hydrolysis of mixed monomolecular films of tricaprylin/dilauroylphosphatidylcholine by lipase and phospholipase A2

Cited by 5 publications

References 19 publications

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

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

CNS myelin structural modification induced in vitro by phospholipases A2

Action of Vipoxin and its separated components on monomolecular film of Dilauroylphosphatidylcholine at the air/water interface

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