Effects of extraction of the H-subunit from Rhodobacter sphaeroides reaction centers on relaxation processes associated with charge separation.

Effects of extraction of the H-subunit from Rhodobacter sphaeroides photosynthetic reaction centers (RC) on the characteristics of the photoinduced conformational transition associated with electron transfer between photoactive bacteriochlorophyll and primary quinone acceptor were studied. Extraction of the H-subunit (i.e., the subunit that is not directly bound to electron transfer cofactors) was found to have a significant effect on the dynamic properties of the protein--pigment complex of the RC, the effect being mediated by modification of parameters of the relaxation processes associated with charge separation.

Dipyridamole and its derivatives modify the kinetics of the electron transport in reaction centers from Rhodobacter sphaeroides.

A well known vasodilator dipyridamole (DIP), 2,6-bis(diethanolamino)-4,8-dipiperidinopyrimido[5,4-d]pyrim idine, and its derivatives have recently been shown as potential co-activators (modulators) in the phenomenon of multidrug resistance (MDR) in cancer therapy. They inhibit the specific function of a transmembrane P-glycoprotein responsible for the ex-flux of anti-cancer drugs from tumor cells. To clarify molecular mechanisms of the anti-MDR activity of DIP and its two derivatives, RA25 and RA47, we have studied their effects on electron transport in reaction centers (RC) from purple photosynthetic bacteria Rb. sphaeroides, using RC as a model system. Increasing concentrations of DIP and RA47 progressively accelerate the back electron transfer from the primary quinone acceptor QA to the bacteriochlorophyll dimer Bchl2 (Bchl2+ -QA- recombination). In the absence of o-phenantroline, when both quinone acceptors QA and QB are involved in the electron transport, RA47 is more effective than DIP. DIP stabilizes the electron on the secondary quinone acceptor QB, the effect manifested as the retardation of Bchl2+ -QB- recombination. Effects of RA25 are negligible in all cases. The drugs are proposed to change the electron transport affecting the RC structural dynamics and the stabilization of the electron on quinone acceptors through modification of H-bonds in the system.

Methods of isolation of reaction center preparations from photosynthetic purple bacteria.

Various modern methods of isolation of functionally active membrane complexes of the reaction centers (RC) from photosynthetic purple bacteria are reviewed. Special attention is given to the methods of RC isolation from bacteria which are widely used in experimental practice. The analysis includes the main steps of RC isolation, evaluation of purity of the resultant preparation, and characterization of its functional activity. Besides description of conventional methods of RC isolation based on ion-exchange chromatography and hydroxyapatite chromatography, some other methods such as affinity chromatography and high-performance chromatography at high and fine pressure are also considered.