Abstract
Using site-directed mutagenesis, we obtained the mutant of the purple bacterium Rhodobacter sphaeroides with Ile to His substitution at position 177 in the L-subunit of the photosynthetic reaction center (RC). The mutant strain forms stable and photochemically active RC complexes. Relative to the wild type RCs, the spectral and photochemical properties of the mutant RC differ significantly in the absorption regions corresponding to the primary donor P and the monomer bacteriochlorophyll (BChl) absorption. It is shown that the RC I(L177)H contains only three BChl molecules compared to four BChl molecules in the wild type RC. Considering the fact that the properties of both isolated and membrane-associated mutant RCs are similar, we conclude that the loss of a BChl molecule from the mutant RC is caused by the introduced mutation but not by the protein purification procedure. The new mutant missing one BChl molecule but still able to perform light-induced reactions forming the charge-separated state P+QA −A appears to be an interesting object to study the mechanisms of the first steps of the primary electron transfer in photosynthesis.
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Abbreviations
- RC:
-
reaction center
- BChl:
-
bacteriochlorophyll
- BPhe a :
-
bacteriopheophytin a
REFERENCES
Mishel, H., Weyer, K. A., Gruenberg, H., Duner, I., Oesterhelt, D., and Lottspeich, F. (1986) EMBO J., 5, 1149–1158.
Maroti, P., Kirmaier, C., Wraight, C., Holten, D., and Pearlstein, R. M. (1985) Biochim. Biophys. Acta, 810, 132–141.
Shuvalov, V. A. (1990) Primary Processes of Light Energy Transformation in Photosynthesis [in Russian], Nauka, Moscow.
Yakovlev, A. G., Vasilieva, L. G., Shkuropatov, A. Ya., Bolgarina, T. I., Shkuropatova, V. A., and Shuvalov, V. A. (2003) J. Phys. Chem. A, 107, 8330–8338.
Bolgarina, T. I., Khatypov, R. A., Vasilieva, L. G., Shkuropatov, A. Ya., and Shuvalov, V. A. (2004) Doklady Ros. Akad. Nauk, 394, 1–4.
Vasilieva, L. G., Bolgarina, T. I., Khatypov, R. A., Shkuropatov, A. Ya., Miyake, J., and Shuvalov, V. A. (2001) Doklady Ros. Akad. Nauk, 376, 826–829.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning–a Laboratory Manual, Cold Spring Harbor, NY.
Simon, R., Priefer, U., and Puhler, A. (1983) Biotechnology, 1, 787–791.
Jones, M. R., Visschers, R. W., van Grondelle, R., and Hunter, C. N. (1992) Biochemistry, 31, 4458–4465.
Keen, N. T., Tamaki, S., Kobayashi, D., and Trollinger, D. (1988) Gene, 70, 191–197.
Ho, S. N., Hunt, H. D., Horton, R. M., Pullen, J. K., and Pease, L. R. (1989) Gene, 77, 51–59.
Shuvalov, V. A., Shkuropatov, A. Ya., Kulakova, S. M., et al. (1986) Biochim. Biophys. Acta, 849, 337–346.
Straley, S. C., Parson, W. W., Mauzerall, D. C., et al. (1973) Biochim. Biophys. Acta, 305, 597–609.
Hammes, S. L., Mazzola, L., Boxer, S. G., Gaul, D. F., and Schenck, C. C. (1990) Proc. Natl. Acad. Sci. USA, 87, 5682–5686.
Jackson, J. A., Lin, S., Taguchi, A. K. W., Williams, J. C., and Woodbury, N. W. (1997) J. Phys. Chem. B, 101, 5747–5754.
Moore, L. J., and Boxer, S. G. (1998) Photosynth. Res., 55, 173–180.
Williams, J. C., Alden, R. G., Murchison, H. A., Peloquin, J. M., Woodbury, N. W., and Allen, J. P. (1992) Biochemistry, 31, 11029–11037.
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Translated from Biokhimiya, Vol. 70, No. 11, 2005, pp. 1527–1533.
Original Russian Text Copyright © 2005 by Khatypov, Vasilieva, Fufina, Bolgarina, Shuvalov.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM05-038, June 5, 2005.
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Khatypov, R.A., Vasilieva, L.G., Fufina, T.Y. et al. Substitution of Isoleucine L177 by Histidine Affects the Pigment Composition and Properties of the Reaction Center of the Purple Bacterium Rhodobacter sphaeroides . Biochemistry (Moscow) 70, 1256–1261 (2005). https://doi.org/10.1007/s10541-005-0256-3
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DOI: https://doi.org/10.1007/s10541-005-0256-3