Protochlorophyllide and protochlorophyll in model membranes — An influence of hydrophobic side chain moiety

Abstract

In the present work, a comparative study of protochlorophyllide- and protochlorophyll-lipid interaction was performed on liposomes prepared from phospholipids and galactolipids, which had a pigment content varying from 0.1 to 4mol%. The incorporation of pigment molecules into the lipid bilayer and pigment–pigment interactions were investigated. Protochlorophyllide entered the lipid bilayer spontaneously and showed fluorescence spectra characteristic of its monomers. Similar spectra were observed for protochlorophyll where its concentration was low. However, the fluorescence maxima of protochlorophyll monomers were blue-shifted compared to those of protochlorophyllide by about 5nm. Protochlorophyll at high concentrations formed transient aggregates that showed an additional fluorescence band with a maximum at around 685nm, especially in liposomes prepared from phospholipids. For both compounds, the Stern–Volmer constant for KI quenching was much lower in liposomes than in solution, which confirmed the incorporation of these compounds into the lipid bilayer. Two populations of protochlorophyll that differed in their accessibility to quenching by KI were determined, and the proportions between them for different lipids are discussed. Protochlorophyllide showed such heterogeneity only in DPPC membranes. Quenching with 5- and 16-SASL revealed a localization of the porphyrin ring of both Pchl and Pchlide in the polar headgroup area of the lipid bilayer. The side chain of protochlorophyll forced these molecules to localize deeper in the bilayer in the case of DPPC in gel phase.