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. 2001 Feb;80(2):749–754. doi: 10.1016/S0006-3495(01)76054-7

Dipalmitoyl-phosphatidylcholine/phospholipase D interactions investigated with polarization-modulated infrared reflection absorption spectroscopy.

I Estrela-Lopis 1, G Brezesinski 1, H Möhwald 1
PMCID: PMC1301273  PMID: 11159442

Abstract

The hydrolysis of 1,2-dipalmitoylphosphatidylcholine (DPPC) catalyzed by Streptomyces chromofuscus phospholipase D (PLD) has been investigated using monolayer techniques and polarization-modulated infrared absorption reflection spectroscopy. The spectroscopic analysis of the phosphate groups provides a quantitative estimation of the hydrolysis yield. The hydrolysis kinetics was investigated in dependence on the phase state of the lipid monolayer. It was found that PLD exhibits maximum activity in the liquid-expanded phase, whereas PLA2 has its activity maximum in the two-phase region. A lag phase was observed in all experiments indicating that small amounts of the hydrolysis product 1,2-dipalmitoylphosphatidic acid (DPPA) are needed for initiating the fast hydrolysis reaction. Higher concentrations of DPPA inhibit the hydrolysis. The critical inhibition concentration of DPPA is a function of the monolayer pressure.

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