Structure and Function of the Vacuolar H+-ATPase: Moving from Low-Resolution Models to High-Resolution Structures
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In the absence of a high-resolution structure for the vacuolar H+-ATPase, a number of approaches can yield valuable information about structure/function relationships in the enzyme. Electron microscopy can provide not only a representation of the overall architecture of the complex, but also a low-resolution map onto which structures solved for individually expressed subunits can be fitted. Here we review the possibilities for electron microscopy of the Saccharomyces V-ATPase and examine the suitability of V-ATPase subunits for expression in high yield prokaryotic systems, a key step towards high-resolution structural studies. We also review the role of experimentally-derived structural models in understanding structure/function relationships in the V-ATPase, with particular reference to the complex of proton-translocating 16 kDa proteolipids in the membrane domain of the V-ATPase. This model in turn makes testable predictions about the sites of binding of bafilomycins and the functional interactions between the proteolipid and the single-copy membrane subunit Vph1p, with implications for the constitution of the proton translocation pathway.
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- Bhandari, P., and Gowrishankar, J. (2003). J. Bacteriol. 179, 4403–4406.Google Scholar
- Chen, J. Q., Acton, T. B., Basu, S. K., Montelione, G. T., and Inouye, M. (2002). J. Mol. Microbiol. Biotech. 4, 519–524.Google Scholar
- Holliday, L. S., Gluck, S. L., and Lee, B. S. (1999). J. Bone Miner. Res. 14, SU195.Google Scholar
- Hunt, I. E., and Bowman, B. J. (1997). J. Bioenerg. Biomemb. 29, 533–540.Google Scholar
- Johnson, W. C., Jr. (1990). Proteins: Struct. Funct. Genet. 7, 205–214Google Scholar
- Jones, D. T., Tress, M., Bryson, K., and Hadley, C. (1999). Proteins: Struct. Funct. Genet. Suppl. 3, 104–111.Google Scholar
- Jones, R. P. O., Hunt, I. E., Jaeger, J., Ward, A., O'Reilly, J., Barratt, E., Findlay, J. B. C., and Harrison, M. A. (2001). Mol. Memb. Biol. 18, 283–290.Google Scholar
- Landolt-Marticorena, C., Williams, K. M., Correa, J., Chen, W., and Manolson, M. F. (2000). J. Biol. Chem. 275, 15449–15457.Google Scholar
- Leng, X. H., Nishi, T., and Forgac, M. (1999). J. Biol. Chem. 274, 14655–14661.Google Scholar
- Powell, B. (1999). PhD Thesis, University of Leeds, Leeds, UK.Google Scholar
- Provencher, S. W. (1982). Comp. Phys. Commun. 27, 229–242.Google Scholar