Gating the selectivity filter in ClC chloride channels

Science

Volumn 300, Issue 5616, 2003, Pages 108-112

  Dutzler, Raimund ^a   Campbell, Ernest B ^a   MacKinnon, Roderick ^a  

^a ROCKEFELLER UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CHLORINE COMPOUNDS; ELECTROLYTES; ESCHERICHIA COLI; IONS;

SELECTIVITY FILTERS;

CELL MEMBRANES;

CARBOXYL GROUP; CHLORIDE CHANNEL; CHLORIDE ION; GLUTAMIC ACID DERIVATIVE; IMMUNOGLOBULIN F(AB) FRAGMENT;

BIOLOGY; CHLORIDE; ELECTRICAL PROPERTY;

ACIDIFICATION; ARTICLE; BINDING SITE; CELL ACTIVITY; CELL MEMBRANE TRANSPORT; CHANNEL GATING; CHLORIDE CONDUCTANCE; CRYSTAL STRUCTURE; ELECTRIC ACTIVITY; ESCHERICHIA COLI; GENE MUTATION; NONHUMAN; PRIORITY JOURNAL; PROTEIN STRUCTURE; TORPEDO;

AMINO ACID SUBSTITUTION; ANIMALS; ANTIBODIES, MONOCLONAL; BINDING SITES; CHLORIDE CHANNELS; CHLORIDES; CRYSTALLOGRAPHY, X-RAY; DIMERIZATION; ESCHERICHIA COLI; ESCHERICHIA COLI PROTEINS; GLUTAMATES; HYDROGEN BONDING; HYDROGEN-ION CONCENTRATION; IMMUNOGLOBULIN FAB FRAGMENTS; ION CHANNEL GATING; MODELS, MOLECULAR; OOCYTES; PATCH-CLAMP TECHNIQUES; POINT MUTATION; PROTEIN CONFORMATION; PROTEIN STRUCTURE, SECONDARY; PROTEIN STRUCTURE, TERTIARY; TORPEDO; XENOPUS;

ESCHERICHIA COLI; TORPEDINIDAE; TORPEDO;

EID: 0037418859     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.1082708     Document Type: Article

References (29)

1. Y. Jiang et al., Nature 417, 523 (2002).
2. D. A. Doyle et al., Science 280, 69 (1998).
3. J. H. Morais-Cabral, Y. Zhou, R, MacKinnon, Nature 414, 37 (2001).
4. Y. Zhou, J. H. Morais-Cabral, A. Kaufman, R. MacKinnon, Nature 414, 43 (2001).
5. E. A. Richard, C. Miller, Science 247, 1208 (1990).
6. M. Maduke, C. Miller, J. A. Mindell, Annu. Rev. Biophys. Biomol. Struct. 29, 411 (2000).
7. T. J. Jentsch, V. Stein, F. Weinreich, A. A. Zdebik, Physiol. Rev. 82, 503 (2002).
8. M. Pusch, U. Ludewig, A. Rehfeldt, T. J. Jentsch, Nature 373, 527 (1995).
9. T. Y. Chen, C. Mitter, J. Gen. Physiol. 108, 237 (1996).
10. R. Dutzler, E. B. Campbell, M. Cadene, B. T. Chait, R. MacKinnon, Nature 415, 287 (2002).
11. 280 ratio of 1:2.3. The complex was purified on a Superdex 200 column (Pharmacia) equilibrated in 150 mM NaCl, 10 mM tris-HCl (pH 7.5), and 10 mM DM. For crystallization, the NaCl concentration was lowered to 90 mM by dilution. For preparation of crystals in NaBr, the protein was dialyzed against a solution containing 90 mM NaBr. Crystals of the E. coli ClC Fab complex were grown in sitting drops at 20°C by equilibrating a 1:1 mixture of protein and reservoir solutions against the reservoir. The reservoir contained 28 to 31% polyethylene glycol (PEG 300), 50 mM glycine (pH 9.5), and 100 mM NaCl. For crystals grown in Br, NaCl was replaced by NaBr. Crystals were frozen in a stream of boiled-off nitrogen. The PEG 300 concentration was raised to 35% by dialysis before freezing. All data sets were collected on frozen crystals.
12. 2, and 100 mM NaCl. For experiments at pH 5.0, Hepes was replaced with 5 mM citrate.
13. C. Miller, Philos. Trans. R. Soc. London Ser. B 299, 401 (1982).
14. T. J. Jentsch, K. Steinmeyer, G. Schwarz, Nature 348, 510 (1990).
15. M. F. Chen, T. Y. Chen, J. Gen. Physiol. 118, 23 (2001).
16. C. Miller, M. M. White, Ann. N.Y. Acad. Sci. 341, 534 (1980).
17. Y, Sun et al., Nature 417, 245 (2002).
18. N. Unwin, A. Miyazawa, J. Li, Y. Fujiyoshi, J. Mol. Biol. 319, 1165 (2002).
19. Y. Jiang et al., Nature 417, 515 (2002).
20. R. Iyer, T. M. Iverson, A, Accardi, C. Miller, Nature 419, 715 (2002).
21. S. Uchida et al., J. Biol. Chem. 268, 3821 (1993).
22. B. Roux, R. MacKinnon, Science 285, 100 (1999).
23. E. Harlow et al., Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989).
24. Z. Otwinowski, W. Minor, Methods Enzymol. 276, 307 (1997).
25. J. Navaza, Acta Crystallogr. A50, 157 (1994).
26. K. Cowtan, Joint CCP4 ESF-EACBM Newsl. Protein Crystalloyr. 31, 34 (1994).
27. T. A. Jones, J. Y. Zou, S. W. Cowan, M. Kjeldgaard, Acta Crystallogr. A47, 110 (1991).
28. A. T. Brünger et al., Acta Crystallogr. D54, 905 (1998).
29. We thank T. Jensch for DNA encoding the ClC-0 channel, the staff at the National Synchrotron Light Source (NSLS) X25, the staff at Cornell High Energy Synchrotron Source (CHESS) A1, M. Zhou and O. Yifrach for assistance with electrophysiology, Y. Zhou for assistance in data collection, members of the MacKinnon laboratory for help at all stages of the project, and T. Y. Chen for manuscript critique. Supported by grants from NIH (R.M.). R.M. is an investigator of the Howard Hughes Medical Institute. Coordinates have been deposited into the Protein Data Bank under codes 10TS, 10TT, and 10TU.