Probing protein structural requirements for formation of the core light-harvesting complex of photosynthetic bacteria using hybrid reconstitution methodology.

The α- and β-polypeptides of LH1 isolated from four different photosynthetic bacteria (Rhodospirillum rubrum, Rhodobacter sphaeroides, Rhodobacter capsulatus and Rhodopseudomonas viridis) were used for homologous and hybrid reconstitution experiments with bacteriochlorophyll a. Formation of B820-type subunit complexes and LH1-type complexes were evaluated. The β-polypeptides of R. rubrum, Rb. sphaeroides and Rb. capsulatus behaved similarly and formed B820-type subunit complexes in the absence of an α-polypeptide. The α- and β-polypeptides were both required to form a LH1-type complex with each of these three homologous systems. In hybrid experiments where the β-polypeptides were tested for reconstitution with α-polypeptides other than their homologous partners, half of the twelve possible combinations resulted in formation of both B820- and LH1-type complexes. Three of the combinations that did not result in formation of a LH1-type complex involved the β-polypeptide of R. rubrum. It is suggested that these latter results can be explained by charge repulsion between the Lys at position-17 (assigning the conserved His located nearest to the C-terminus as position 0) in the β-polypeptide of R. rubrum and each of the heterologous α-polypeptides tested, all of which have an Arg at this location. Conclusions that can be derived from these experimental results include: (1) the experimental data support the idea that a central core region of approximately 40 amino acids exists in each of the polypeptides, which contains sufficient information to allow formation of both the B820- and LH1-type complexes and (2) a specific portion of the N-terminal hydrophilic region of each polypeptide was found in which ion pairs between oppositely charged groups on the α- and β-polypeptides seem to stabilize complex formation.