X-ray Structure Determination of the Cytochrome c2: Reaction Center Electron Transfer Complex from Rhodobacter sphaeroides

Abstract

In the photosynthetic bacterium Rhodobacter sphaeroides, a water soluble cytochrome c 2 (cyt c 2) is the electron donor to the reaction center (RC), the membrane-bound pigment–protein complex that is the site of the primary light-induced electron transfer. To determine the interactions important for docking and electron transfer within the transiently bound complex of the two proteins, RC and cyt c 2 were co-crystallized in two monoclinic crystal forms. Cyt c 2 reduces the photo-oxidized RC donor (D +), a bacteriochlorophyll dimer, in the co-crystals in ∼0.9 μs, which is the same time as measured in solution. This provides strong evidence that the structure of the complex in the region of electron transfer is the same in the crystal and in solution. X-ray diffraction data were collected from co-crystals to a maximum resolution of 2.40 Å and refined to an R-factor of 22% ( R free=26%). The structure shows the cyt c 2 to be positioned at the center of the periplasmic surface of the RC, with the heme edge located above the bacteriochlorophyll dimer. The distance between the closest atoms of the two cofactors is 8.4 Å. The side-chain of Tyr L162 makes van der Waals contacts with both cofactors along the shortest intermolecular electron transfer pathway. The binding interface can be divided into two domains: (i) A short-range interaction domain that includes Tyr L162, and groups exhibiting non-polar interactions, hydrogen bonding, and a cation–π interaction. This domain contributes to the strength and specificity of cyt c 2 binding. (ii) A long-range, electrostatic interaction domain that contains solvated complementary charges on the RC and cyt c 2. This domain, in addition to contributing to the binding, may help steer the unbound proteins toward the right conformation.