Protochlorophyllide in model systems - An approach to in vivo conditions

Absorption and fluorescence properties of protochlorophyllide (Pchlide) monomers and aggregates in various model systems are presented in this study. The absorption and fluorescence maxima, and fluorescence lifetimes of Pchlide monomers were not dependent on liposome composition. Fluorescence quenching experiments using KI and SASLs as fluorescence quenchers, revealed that Pchlide molecules entered a lipid bilayer and were localized close to the polar lipid headgroup area. The process of Pchlide aggregation was evident for high (i.e. at least 9 mol%) Pchlide content in liposomes prepared from galactolipids. To our knowledge, this is the first study of Pchlide aggregation in membrane-mimicking model systems. The aggregates showed absorption maxima at 480 and 650 nm. Fluorescence of the aggregates measured for excitation at 480 nm had a maximum at 656 nm and was characterized with two fluorescence lifetime components, i.e. 0.1 and 1-2 ns. Pchlide aggregates observed in the buffer had similar position of absorption and fluorescence bands to those observed in liposomes, although the overall fluorescence intensity was considerably lower. Some differences in the relative intensity of Soret absorption bands were observed. These results showed that the presence of liposomes decreased the efficiency of the process of Pchlide aggregation. Water bound at the interface region of AOT/isooctane/water reversed micelles induced disaggregation of the Pchlide aggregates indicating that Pchlide aggregates are buried into hydrophilic core of micelles. The results are discussed with respect to the role of lipids in Pchlide aggregation found in plant etioplasts and their significance for light-induced Pchlide photoreduction.

Highlights► Absorption and fluorescence of protochlorophyllide was studied in model systems. ► Galactolipids facilitate protochlorophyllide aggregation. ► Fluorescence lifetimes of Pchlide aggregates were 0.1 and 1.5-2 ns. ► Fluorescence lifetimes of Pchlide monomers in liposomes were of 4.1-4.6 ns. ► Molecular dynamics of water is important for Pchlide aggregation.