Topological study of PSI‐A and PSI‐B, the large subunits of the photosystem‐I reaction center

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Abstract
The core of the photosystem-I reaction center is formed by polypeptides PSI-A and PSI-B, the products of the homologous psaA and psaB genes. Based on hydropathy analyses, models have been proposed for the folding of these polypeptide chains in the membrane [Fish, L. E., Kück, U. & Bogorad, L. (1985), in Molecular biology of the photosynthetic apparatus, pp. 111-120, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY]. To test these models, we have tried to identify regions of PSI-A that are exposed to the surrounding medium, on the stromal or lumenal surface of the membrane. Immunogold labeling of thylakoid vesicles, with antibodies to synthetic peptides, shows that residues 413-421 of PSI-A are exposed on the stromal surface of the membrane, and that the accessibility of this region is enhanced by NaSCN treatment, which removes extrinsic polypeptides. This treatment also enhances a trypsin-cleavage site which may lie just after residues 413-421. Immunogold labeling also indicates that residues 371-379 and 497-505 are exposed on the lumenal surface. These results establish the conformation of the central portion of the polypeptide. Assuming that the transmembrane regions are correctly predicted by the 11-helix model, the N-terminal domain, as well as the conserved region proposed to bind the iron-sulfur center FX, would be expected to be on the stromal surface.