Structure and properties in solution of PsaD, an extrinsic polypeptide of photosystem I.
PsaD is a small, extrinsic polypeptide located on the stromal side (cytoplasmic side in cyanobacteria) of the photosystem I reaction centre complex. The gene from the cyanobacterium Nostoc sp. PCC 8009 was expressed in Escherichia coli and the structure of the recovered protein in solution investigated. Size-exclusion chromatography, dynamic light scattering and measurement of 15N transverse relaxation times showed that the protein is a stable dimer in solution, whereas in the reaction centre complex it is a monomer. NMR experiments showed that the dimer is symmetrical and that there are at least two domains, one structured and the remainder unstructured. The structured domain contains a small amount of beta-sheet. Three-dimensional heteronuclear NMR spectra of [13C, 15N]PsaD showed that the structured domain is associated with the central part of the sequence while the N- and C-terminal regions are mobile. Evidence was obtained for a shift in equilibrium between two slightly different conformational states at about pH 6, and the protein was shown to bind to PsaE preferentially at neutral pH. Addition of trifluoroethanol was shown to induce the formation of a small amount of alpha-helix, and the form present in 30% trifluoroethanol appears to be more closely related to the in situ structure, which has been reported to contain one short helix in crystals [Schubert, W.-D., Klukas, O., Krauss, N., Saenger, W., Fromme, P. & Witt, H. T. (1997) J. Mol. Biol. 272, 741-769]. The significance of these findings for the assembly of the complex is discussed.
Department of Biochemistry and Centre for Molecular Recognition, University of Cambridge, England.
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Xia
R W
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Broadhurst
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Laue
D A
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Bryant
J H
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Golbeck
D S
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Bendall
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D000458:Cyanobacteria; D019281:Dimerization; D008027:Light; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D045331:Photosystem I Protein Complex; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D012996:Solutions
D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D010940:Plant Proteins; C070290:PsaD protein, cyanobacteria; D011994:Recombinant Proteins; D012996:Solutions; C079741:photosystem I, psaE subunit