Abstract
We have isolated the water-soluble BChla-protein (FMO-protein) from the greer sulfur bacteriumChlorobium tepidum by a new procedure involving a salt-wash of isolated membranes at alkaline pH. The absorption spectrum of the isolated FMO-protein at 77 K was compared with that of a reaction-center complex containing the FMO-protein (FMO-RC complex) isolated fromC. tepidum following the procedure of Feiler U, Nitsche W and Michel H (1992) Biochemistry 31: 2608–2614. Oxidation or illumination of the FMO-RC complex caused bleaching of a component with a maximum at 836 nm which was not present in the purified FMO-protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BChl:
-
bacteriochlorophyll
- CD:
-
circular dichroism
- FMO:
-
Fenna-Matthews-Olson
- RC:
-
reaction center
- TMBZ:
-
3,3′,5,5′-tetramethylbenzidine
References
Buettner M, Xie D-L, Nelson H, Pinther W, Hauska G and Nelson N (1992) Photosynthetic reaction center genes in green sulfur bacteria and in Photosystem 1 are related. Proc Natl Acad Sci USA 89: 8135–8139
Dracheva S, Williams JC and Blankenship RE (1992) Cloning and sequencing of the FMO-protein gene fromChlorobium tepidum. In: Murata N (ed) Research in Photosynthesis Vol I, pp 53–56. Kluwer Academic Publishers, Dordrecht
Feiler U, Nitschke W and Michel H (1992) Characterization of an improved reaction center preparation from the photosynthetic green sulfur bacteriumChlorobium containing the FeS centers FA and FB and a bound cytochrome subunit. Biochemistry 31: 2608–2614
Fling SP and Gregerson DS (1986) Peptide and protein molecular weight determination by electrophoresis using a high-molarity Tris buffer system without urea. Anal Biochem 155: 83–88
Hurt EC and Hauska G (1984) Purification of membrane-bound cytochromes and a photoactive P840 protein complex of the green sulfur bacteriumChlorobium limicola f.thiosulfatophilum. FEBS Lett 168: 149–154
Illinger N, Xie D-L, Hauska G and Nelson N (1993) Identification of the subunit carrying FeS-centersA andB in the P840-reaction center preparation ofChlorobium limicola. Photosynth Res 38: 111–114
Kramer HJM, Kingma H, Swarthoff T and Amesz J (1982) Prompt and delayed fluorescence in pigment-protein complexes of a green photosynthetic bacterium. Biochim Biophys Acta 681: 359–364
Matthews BW, Fenna RE, Bolognesi MC, Schmid MF and Olson JM (1979) Structure of a bacteriochlorophylla-protein from the green photosynthetic bacteriumProsthecochloris aestuarii. J Mol Biol 131: 259–285
Miller M, Liu X, Snyder SW, Thurnauer MC and Biggins J (1992) Photosynthetic electron-transfer reactions in the green sulfur bacteriumChlorobium vibrioforme: Evidence for the functional involvement of iron-sulfur redox centers on the acceptor side of the reaction center. Biochemistry 31: 4354–4363
Oh-oka H, Kakutani S, Matsubara H, Malkin R and Itoh S (1993) Isolation of the photoactive reaction center complex that contains three types of Fe-S centers and a cytochromec subunit from the green sulfur bacteriumChlorobium limicola f.thiosulfatophilum, strain Larsen. Plant Cell Physiol 34: 93–101
Okkels JS, Kjær B, Hansson O, Svendsen I, Møller BL and Scheller HV (1992) A membrane-bound monoheme cytochromec 551 of a novel type is the immediate electron donor to P840 of theChlorobium vibrioforme photosynthetic reaction center complex. J Biol Chem 267: 21139–21145
Olson JM Ke B and Thompson KH (1976) Exciton interaction among chlorophyll molecules in bacteriochlorophylla proteins and bacteriochlorophylla reaction center complexes from green bacteria. Biochim Biophys Acta 430: 524–537
Olson JM, Prince RC and Brune DC (1977) Reaction-center complexes from green bacteria. Brookhaven Symposia in Biology 28: 238–245
Olson JM (1978) Bacteriochlorophylla-proteins from green bacteria. In: Clayton RK and Sistrom WR (eds) The Photosynthetic Bacteria, pp 161–178. Plenum, New York
Olson JM (1980) Chlorophyll organization in green photosynthetic bacteria. Biochim Biophys Acta 594: 33–51
Otte SCM, van derHeiden JC, Pfennig N and Amesz J (1991) A comparative study of the optical characteristics of intact cells of photosynthetic green sulfur bacteria containing bacteriochlorophyllc, d ore. Photosynth Res 28: 77–87
Savitsky A and Golay MJE (1964) Smoothing and differentiation of data by simplified least-squares procedures. Analyt Chem 36: 1627–1639
Swarthoff T and Amesz J (1979) Photochemically active pigment-protein complexes from the green photosynthetic bacteriumProsthecochloris aestuarii. Biochim Biophys Acta 548: 427–432
Sybesma C and Olson JM (1963) Transfer of chlorophyll excitation energy in green photosynthetic bacteria. Proc Natl Acad Sci USA 49: 248–253
Thomas PE, Ryan D and Lewin W (1976) An improved staining procedure for the detection of the peroxidase activity of cytochrome P-450 on sodium dodecyl sulfate polyacrylamide gels. Anal Biochem 75: 168–176
Wahlund TM, Woese CR, Castenholz RW and Madigan M (1991) A thermophilic green sulfur bacterium from New Zealand hot springs,Chlorobium tepidum sp. nov. Arch Microbiol 156: 81–90
Vasmel H, Swarthoff T, Kramer HJM and Amesz J (1983) Isolation and properties of a pigment-protein complex associated with the reaction center of the green photosynthetic sulfur bacteriumProsthecochloris aestuarii. Biochim Biophys Acta 725: 361–367
Whitten WB, Pearlstein RM and Olson JM (1979) New spectral components in high resolution absorption spectra of green bacterial reaction center complexes at 5 K. Photochem Photobiol 29: 823–828
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miller, M., Cox, R.P. & Olson, J.M. Low-temperature spectroscopy of isolated FMO-protein and a membrane-free reaction center complex from the green sulfur bacteriumChlorobium tepidum . Photosynth Res 41, 97–103 (1994). https://doi.org/10.1007/BF02184149
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02184149