Abstract
The bacterial histidine permease, an ABC transporter, from Salmonella typhimurium is composed of a membrane-bound complex, HisQMP2, comprising two hydrophobic subunits (HisQ and HisM), two copies of an ATP-hydrolyzing subunit, HisP, and a soluble receptor, HisJ. We describe the purification and characterization of HisQMP2 using a 6-histidines extension at the carboxy terminus of HisP [HisQMP2(his6)]. The purification is rapid and effective, giving a seven–fold purification with a yield of 85 and 98% purity. Two procedures are described differing in the detergent used (decanoylsucrose and octylglucoside, respectively) and in the presence of phospholipid. HisQMP2(his6) has ATPase and transport activities upon reconstitution into proteoliposomes (PLS). HisQMP2(his6) has a low level ATPase activity (intrinsic activity), which is stimulated to a different extent by the receptor—liganded and unliganded. Its pH optimum is 7.8–8.0, it requires a cation for activity and it displays cooperativity for ATP. The effect of various ATP analogs was analyzed. Determination of the molecular size of HisQMP2(his6) indicates that it is a monomer. The permeability properties of two kinds of reconstituted PLS preparations are described.
Similar content being viewed by others
REFERENCES
Ames, G. F.-L. (1990). In Bacterial Energetics (Krulwich, T. A., ed.) Academic Press, New York, pp. 225–245.
Ames, G. F.-L., Mimura, C., Holbrook, S., and Shyamala, V. (1992). Advan. Enzymol. 65, 1–47.
Ames, G. F.-L., Liu, C. E., Joshi, A. K., and Nikaido, K. (1996). J. Biol. Chem. 271, 14264–14270.
Barela, T. D. and Sherry, A. D. (1976). Anal. Biochem. 71, 351–357.
Chifflet, S., Torriglia, A., Chiesa, R., and Tolosa, S. (1988). Anal. Biochem. 168, 1–4.
Davidson, A. L. and Nikaido, H. (1991). J. Biol. Chem. 266, 8946–8951.
Doige, C. A. and Ames, G. F.-L. (1993). Annu. Rev. Microbiol. 47, 291–319.
Higgins, C. F. (1992). Annu. Rev. Cell Biol. 8, 67–113.
Higgins, C. F. and Ames, G. F.-L. (1981). Proc. Natl. Acad. Sci. USA 78, 6038–6042.
Hobson, A., Weatherwax, R., and Ames, G. F.-L. (1984). Proc. Natl. Acad. Sci. USA 81, 7333–7337.
Hung, L.-W., Wang, I. X., Nikaido, K., Liu, P.-Q., Ames, G. F.-L., and Kim, S.-H. (1998). Nature (London) 396, 703–707.
Hyde, S. C., Emsley, P., Hartshorn, M. J., Mimmack, M. M., Gileadi, U., Pearce, S. R., Gallagher, M. P., Gill, D. R., Hubbard, R. E., and Higgins, C. F. (1990). Nature (London) 346, 362–365.
Kerppola, R. E. and Ames, G. F.-L. (1992). J. Biol. Chem. 267, 2329–2336.
Kerppola, R. E., Shyamala, V., Klebba, P., and Ames, G. F.-L. (1991). J. Biol. Chem. 266, 9857–9865.
Kreimer, D. I., Chai, K. P., and Ames, G. F.-L. (2000). Biochemistry 39, 14183–14195.
Kustu, S. G. and Ames, G. F.-L. (1973). J. Bacteriol. 116, 107–113.
Lill and Wickner, W. (1990). Cell 60, 271–280.
Liu, C. E. (1996). Ph.D. Thesis. University of California at Berkeley, Berkeley, California.
Liu, C. E. and Ames, G. F.-L. (1997). J. Biol. Chem. 272, 859–866.
Liu, C. E., Liu, P.-Q., and Ames, G. F.-L. (1997). J. Biol. Chem. 272, 21883–21891.
Liu, P.-Q. (1997). Ph.D. Thesis. University of California at Berkeley, Berkeley, California.
Liu, P.-Q. and Ames, G. F.-L. (1998). Proc. Natl. Acad. Sci. USA 95, 3495–3500.
Liu, P.-Q., Liu, C. E., and Ames, G. F.-L. (1999). J. Biol. Chem. 274, 18310–18318.
Martin, R. G. and Ames, B. N. (1961). J. Biol. Chem. 236, 1372–1379.
Mimura, C. S., Admon, A., Hurt, K. A., and Ames, G. F.-L. (1990).J. Biol. Chem. 265, 19535–19542.
Mimura, C. S., Holbrook, S. R., and Ames, G. F.-L. (1991). Proc. Natl. Acad. Sci. USA 88, 84–88.
Nikaido, H. (1994). FEBS Lett. 346, 55–58.
Nikaido, K. and Ames, G. F.-L. (1992). J. Biol. Chem. 267, 20706–20712.
Nikaido, K. and Ames, G. F.-L. (1999). J. Biol. Chem. 274, 26727–26735.
Nikaido, K., Liu, P.-Q., and Ames, G. F.-L. (1997). J. Biol. Chem. 272, 27745–27752.
Peterson, G. L. (1977). Anal. Biochem. 83, 346–356.
Petronilli, V. and Ames, G. F.-L. (1991). J. Biol. Chem. 266, 16293–16296.
Sharom, F. J., Liu, R., Romsicki,Y., and Lu, P. (1999). Biochim. Biophys. Acta 1461, 327–345.
Shyamala, V., Baichwal, V., Beall, E., and Ames, G. F.-L. (1991). J. Biol. Chem. 266, 18714–18719.
Strauss, E. (1999). Science 285, 814–815.
Wolf, A., Shaw, E. W., Oh, B.-H., De Bondt, H., Joshi, A. K., and Ames, G. F.-L. (1995). J. Biol. Chem. 270, 16097–16106.
Young, J. and Holland, I. B. (1999). Biochim. Biophys. Acta 1461, 177–200.
Young, S. G. and Fielding, C. J. (1999). Nature Genet. 22, 316–318.
Rights and permissions
About this article
Cite this article
Ames, G.FL., Nikaido, K., Wang, I.X. et al. Purification and Characterization of the Membrane-Bound Complex of an ABC Transporter, the Histidine Permease. J Bioenerg Biomembr 33, 79–92 (2001). https://doi.org/10.1023/A:1010797029183
Issue Date:
DOI: https://doi.org/10.1023/A:1010797029183