Abstract
The response of V1 ATPase of the tobacco hornwormManduca sexta to Mg2+ and nucleotide binding in the presence of the enhancer methanol has been studied by CuCl2-induced disulfide formation, fluorescence spectroscopy, and small-angle X-ray scattering. When the V1 complex was supplemented with CuCl2 nucleotide-dependence of A-B-E and A-B-E-D cross-linking products was observed in absence of nucleotides and presence of MgADP+Pi but not when MgAMP·PNP or MgADP were added. A zero-length cross-linking product of subunits D and E was formed, supporting their close proximity in the V1 complex. The catalytic subunit A was reacted with N-4[4-[7-(dimethylamino)-4-methyl]coumarin-3-yl]maleimide (CM) and spectral shifts and changes in fluorescence intensity were detected upon addition of MgAMP·PNP, -ATP, -ADP+Pi, or -ADP. Differences in the fluorescence emission of these nucleotide-binding states were monitored using the intrinsic tryptophan fluorescence. The structural composition of the V1 ATPase from M. sexta and conformational alterations in this enzyme due to Mg2+ and nucleotide binding are discussed on the basis of these and previous observations.
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REFERENCES
Adachi, I., Puopolo, K., Marquez-Sterling, N., Arai, H., and Forgac, M. (1990). J. Biol. Chem. 265, 967-973.
Aviezer-Hagai, K., Padler-Karavani, V., and Nelson, N. (2003). J. Exp. Biol. 206, 3227-3237.
Boulin, C., Kempf, R., Koch, M. H. J., and McLaughlin, S. M. (1986). Nucl. Instrum. Methods A 249, 399-407.
Bowman, B. J., and Bowman, E. J. (1997). In The Mycota III. Biochemistry And Molecular Biology (Brambl, R., and Marzluf, G. A. eds.)., pp. 57-83.
Coskun, ü., Grüber, G., Koch, M. H. J., Godovac-Zimmermann, J., Lemker, T., and Müller, V. (2002). J. Biol. Chem. 277, 17327-17333.
Damerval, C., le Guillioux, M., Blaisomeau, J., and de Vienne, D. (1987). Electrophoresis 8, 158-159.
Flores, G. O., Acosta, A., and Gomeź Puyou, A. (1982). Biochim. Biophys. Acta 679, 466-473.
Futai, M., Oka, T., Sun-Wada, G.-H., Moriyama, Y., Kanazawa, H., and Wada, Y. (2000). J. Exp. Biol. 203, 107-116.
Gabriel, A., and Dauvergne, F. (1982). Nucl. Instrum. Methods 201, 223-224.
Gibbons, C., Montgomery, M. G., Leslie, A. G., and Walker, J. E. (2000). Nat. Struct. Biol. 7, 1055-1061.
Gräf, R., Harvey, W. R., and Wieczorek, H. (1996). J. Biol. Chem. 271, 20908-20913.
Gräf, R., Novak, F. J. S., Harvey, W. R., and Wieczorek, H. (1992). FEBS Lett. 300, 119-122.
Grüber, G., (2000). J. Bioenerg. Biomembr. 32, 341-346.
Grüber, G., Radermacher, M., Ruiz, T., Godovac-Zimmermann, J., Canas, B., Kleine-Kohlbrecher, D., Huss, M., Harvey, W. R., and Wieczorek, H. (2000a). Biochemistry 39, 8609-8616.
Grüber, G., Svergun, D. I., Coskun, ü., Lemker, T., Koch, M. H. J., Schägger, H., and Müller, V. (2000b). Biochemistry 40, 1890-1896.
Hicks, D. B., and Krulwich, T. A. (1986). J. Biol. Chem. 261, 12896-12902.
Hilario, E., and Gogarten, J. P. (1998). J. Mol. Evol. 46, 703-715.
Hirata, T., Iwamoto-Kihara, A., Sun-Wada, G.-H., Okajima, T., Wada, Y., and Futai, M. (2003). J. Biol. Chem. 278, 23714-23719.
Ihara, K., Abe, T., Sugimura, K. I., and Mukohata, Y. (1992). J. Exp. Biol. 172, 475-485.
Imamura, H., Nakano, M., Nioji, H., Muneyuki, E., Ohkuma, S., Yoshida, M., and Yokoyama, K. (2003). Proc. Natl. Acad. Sci. U.S.A. 100, 2312-2315.
Ito, N., Phillips, S. E. V., Stevens, C., Ogel, Z. B., McPherson, M. J., Keen, J. N., Yadav, K. D. S., and Kowles, P. F. (1991). Nature 350, 87-90
Kane, P. M. (2000). FEBS Lett. 469, 137-141.
Koch, M. H. J., and Bordas, J. (1983). Nucl. Instrum. Methods 208, 461-469.
Laemmli, U. K. (1970). Nature 227, 680-685.
Lerch, K. (1982). J. Biol. Chem. 257, 6414-6419
McCarty, R. E., and Racker, E. (1968). J. Biol. Chem. 243, 129-137.
Merzendorfer, H., Reineke, S., Zhao, X.-F., Jakobmeier, B., Harvey, W. R., and Wieczorek, H. (2000). Biochim. Biophys. Acta 1467, 369-379.
Müller, V., and Grüber, G. (2003). Cell Mol. Life Sci. 60, 474-494.
Nishi, T., and Forgac, M. (2002). Nat. Rev. Mol. Cell. Biol. 3, 94-103.
Novak, F. J. S., Gräf, R., Waring, R., Wolfersberger, M. G., Wieczorek, H., and Harvey, W. R. (1992). Biochim. Biophys. Acta 1132, 67-71.
Pedersen, P. L., Ko, Y. H., and Hong, S. (2000). J. Bioenerg. Biomembr. 32, 325-332.
Radermacher, M., Ruiz, T., Wieczorek, H., and Grüber, G. (2001). J. Struct. Biol. 135, 26-37.
Rizzo, V. F., Coskun, ü., Radermacher, M., Ruiz, T., Armbrüster, A., and Grüber, G. (2003). J. Biol. Chem. 278, 270-275.
Schuster, S. M. (1979). Biochemistry 18, 1162-1167.
Svergun, D. I., Konrad, S., Huss, M., Koch, M. H. J., Wieczorek, H., Altendorf, K., Volkov, V. V., and Grüber, G. (1998). Biochemistry 37, 17659-17663.
Svergun, S. I. (1992). J. Appl. Crystallogr. 25, 495-503.
Xu, T., Vasilyeva, E., and Forgac, M. (1999). J. Biol. Chem. 274, 28909-28915.
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Coskun, Ü., Rizzo, V.F., Koch, M.H.J. et al. Ligand-Dependent Structural Changes in the V1 ATPase from Manduca sexta . J Bioenerg Biomembr 36, 249–256 (2004). https://doi.org/10.1023/B:JOBB.0000031976.44466.6e
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DOI: https://doi.org/10.1023/B:JOBB.0000031976.44466.6e