{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T03:11:15Z","timestamp":1792552275139,"version":"4.5.0"},"reference-count":35,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5360","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1998,4,3]]},"abstract":"<jats:p>\n                    The potassium channel from\n                    <jats:italic>Streptomyces lividans<\/jats:italic>\n                    is an integral membrane protein with sequence similarity to all known K\n                    <jats:sup>+<\/jats:sup>\n                    channels, particularly in the pore region. X-ray analysis with data to 3.2 angstroms reveals that four identical subunits create an inverted teepee, or cone, cradling the selectivity filter of the pore in its outer end. The narrow selectivity filter is only 12 angstroms long, whereas the remainder of the pore is wider and lined with hydrophobic amino acids. A large water-filled cavity and helix dipoles are positioned so as to overcome electrostatic destabilization of an ion in the pore at the center of the bilayer. Main chain carbonyl oxygen atoms from the K\n                    <jats:sup>+<\/jats:sup>\n                    channel signature sequence line the selectivity filter, which is held open by structural constraints to coordinate K\n                    <jats:sup>+<\/jats:sup>\n                    ions but not smaller Na\n                    <jats:sup>+<\/jats:sup>\n                    ions. The selectivity filter contains two K\n                    <jats:sup>+<\/jats:sup>\n                    ions about 7.5 angstroms apart. This configuration promotes ion conduction by exploiting electrostatic repulsive forces to overcome attractive forces between K\n                    <jats:sup>+<\/jats:sup>\n                    ions and the selectivity filter. The architecture of the pore establishes the physical principles underlying selective K\n                    <jats:sup>+<\/jats:sup>\n                    conduction.\n                  <\/jats:p>","DOI":"10.1126\/science.280.5360.69","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:42:47Z","timestamp":1027748567000},"page":"69-77","source":"Crossref","is-referenced-by-count":6198,"title":["The Structure of the Potassium Channel: Molecular Basis of K\n                    <sup>+<\/sup>\n                    Conduction and Selectivity"],"prefix":"10.1126","volume":"280","author":[{"given":"Declan A.","family":"Doyle","sequence":"first","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joa\u0303o Morais","family":"Cabral","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard A.","family":"Pfuetzner","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anling","family":"Kuo","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacqueline M.","family":"Gulbis","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven L.","family":"Cohen","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian T.","family":"Chait","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roderick","family":"MacKinnon","sequence":"additional","affiliation":[{"name":"D. A. Doyle, R. A. Pfuetzner, A. Kuo, and R. MacKinnon are in the Laboratory of Molecular Neurobiology and Biophysics and the Howard Hughes Medical Institute, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. J. M. Cabral and J. M. Gulbis are in the Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. S. L. Cohen and B. T. Chait are in the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller..."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","unstructured":"B. Hille Ionic Channels of Excitable Membranes (Sinauer Sunderland MA ed. 2 1992)."},{"key":"e_1_3_1_3_2","first-page":"409","volume":"72","author":"Hille B.","year":"1978","unstructured":"A. L. Hodgkin, R. D. Keynes, J. Physiol. (London) 128, 61 (1955); S. Hagiwara, S. Miyazaki, S. Krasne, S. Ciani, J. Gen. Physiol. 70, 269 (1977); Hille B., Schwartz W., ibid 72, 409 (1978); J. Neyton and C. Miller, ibid. 92, 549 (1988).","journal-title":"ibid"},{"key":"e_1_3_1_4_2","first-page":"491","volume":"50","author":"Armstrong C. M.","year":"1966","unstructured":"C. M. Armstrong, L. Binstock, J. Gen. Physiol. 48, 859 (1965); Armstrong C. M., ibid 50, 491 (1966); ibid. 54, 553 (1969);ibid. 58, 413 (1971).","journal-title":"ibid"},{"key":"e_1_3_1_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1279807"},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","first-page":"10335","DOI":"10.1021\/bi970988i","volume":"36","author":"Heginbothan L.","year":"1997","unstructured":"H. Schempf, et al., EMBO J. 14, 5170 (1995); Heginbothan L., Odessey E., Miller C., Biochemistry 36, 10335 (1997); D. Marien Cortes and E. Perozo,ibid., p. 10343.","journal-title":"Biochemistry"},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1038\/350232a0","volume":"350","author":"MacKinnon R.","year":"1991","unstructured":"MacKinnon R., Nature350, 232 (1991).","journal-title":"Nature"},{"key":"e_1_3_1_8_2","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1038\/376690a0","volume":"376","author":"Ketchum K. A.","year":"1995","unstructured":"Certain K+ channels contain the equivalent of two subunits in a single open reading frame. These are thought to form the tetramer through the assembly of two dimer subunits [Ketchum K. A., et al., Nature 376, 690 (1995)].","journal-title":"Nature"},{"key":"e_1_3_1_9_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.280.5360.106"},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"1557","DOI":"10.1016\/S0969-2126(97)00305-5","volume":"5","author":"Kleywegt G. J.","year":"1998","unstructured":"Kleywegt G. J., Read R. J., Structure5, 1557 (1998).","journal-title":"Structure"},{"key":"e_1_3_1_11_2","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1006\/jmbi.1994.1690","volume":"243","author":"Kreusch A.","year":"1994","unstructured":"J. Deisenhofer, et al., Nature 318, 618 (1985); S. W. Cowan, et al., ibid 358, 727 (1992); Kreusch A., Schulz G. E., J. Mol. Biol. 243, 891 (1994).","journal-title":"J. Mol. Biol."},{"key":"e_1_3_1_12_2","doi-asserted-by":"crossref","first-page":"1382","DOI":"10.1126\/science.2476850","volume":"245","author":"MacKinnon R.","year":"1989","unstructured":"MacKinnon R., Miller C., Science245, 1382 (1989).","journal-title":"Science"},{"key":"e_1_3_1_13_2","first-page":"123","volume":"16","author":"Naranjo D.","year":"1996","unstructured":"R. MacKinnon, L. Heginbotham, T. Abramson, Neuron 5, 767 (1990); M. Stocker, C. Miller, Proc. Natl. Acad. Sci. U.S.A. 91, 9509 (1994); S. A. N. Goldstein, D. J. Pheasant, C. Miller, Neuron 12, 1377 (1994); P. Hidalgo, R. MacKinnon, Science 268, 307 (1995); J. Aiyar, et al., Neuron 15, 1169 (1995); Naranjo D., Miller C., ibid 16, 123 (1996); R. Ranganathan, J. H. Lewis, R. MacKinnon,ibid., p. 131; A. Gross and R. MacKinnon,ibid., p. 399.","journal-title":"ibid"},{"key":"e_1_3_1_14_2","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1085\/jgp.59.4.388","volume":"59","author":"Armstrong C. M.","year":"1972","unstructured":"Armstrong C. M., Hille B., J. Gen. Physiol.59, 388 (1972).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_15_2","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1126\/science.2218530","volume":"250","author":"MacKinnon R.","year":"1990","unstructured":"MacKinnon R., Yellen G., Science250, 276 (1990).","journal-title":"Science"},{"key":"e_1_3_1_16_2","first-page":"939","volume":"251","author":"Yellen G.","year":"1991","unstructured":"Yellen G., Jurman M. E., Abramson T., MacKinnon R., ibid251, 939 (1991).","journal-title":"ibid"},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80357-8"},{"key":"e_1_3_1_18_2","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/j.1749-6632.1975.tb31481.x","volume":"264","author":"Parsegian V. A.","year":"1975","unstructured":"Parsegian V. A., Ann. N.Y. Acad. Sci.264, 161 (1975).","journal-title":"Ann. N.Y. Acad. Sci."},{"key":"e_1_3_1_19_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1502559"},{"key":"e_1_3_1_20_2","unstructured":"The temperature factors for Val 76 and Gly 77 main chain atoms (but not side chain atoms) refined to higher values than for neighboring atoms. This result can be explained by the difference Fourier analysis which shows alternative positions of the inner K + ion in the selectivity filter and therefore by inference alternative conformations of the coordinating main chain atoms depending on the location of the K + ion."},{"key":"e_1_3_1_21_2","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1085\/jgp.60.5.588","volume":"60","author":"Bezanilla F.","year":"1972","unstructured":"Bezanilla F., Armstrong C. M., J. Gen. Physiol.60, 588 (1972).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_22_2","first-page":"669","volume":"61","author":"Hille B.","year":"1973","unstructured":"Hille B., ibid61, 669 (1973).","journal-title":"ibid"},{"key":"e_1_3_1_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/309453a0"},{"key":"e_1_3_1_24_2","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1085\/jgp.92.5.569","volume":"92","author":"Neyton J.","year":"1988","unstructured":"Neyton J., Miller C., J. Gen. Physiol.92, 569 (1988).","journal-title":"J. Gen. Physiol."},{"key":"e_1_3_1_25_2","unstructured":"The KcsA gene was subcloned into pQE60 (Qiagen) vector and expressed in E. coli XL-1 Blue cells upon induction with 1-\u03b2- d -thiogalactopyranoside. The carboxyl-terminal histidine tagged protein was extracted by homogenization and solubilization in 40 mM decylmaltoside (Anatrace). The KcsA channel was purified on a cobalt affinity column. Thirty-five carboxyl terminal amino acids were cleaved by chymotrypsin proteolysis. The truncated channel was purified to homogeneity by gel filtration and the detergent exchanged in a final dialysis step against 5 mM N N-dimethyldodecylamine-N-oxide (LDAO). Crystals were grown at 20\u00b0C with the sitting drop method by mixing equal volumes of protein solution [5 to 10 mg\/ml 150 mM KCl 50 mM tris (pH 7.5) and 2 mM dithiothreitol] with reservoir mixture [200 mM CaCl 2 100 mM Hepes (pH 7.5) and 48% PEG 400]. Through the entire preparation the channel protein was maintained in solutions containing 150 mM KCl. For definition of K + sites crystals were transferred into solutions where 150 mM KCl was replaced by 150 mM RbCl or 150 mM CsCl."},{"key":"e_1_3_1_26_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1063\/1.1147503","volume":"67","author":"Thiel D.J.","year":"1996","unstructured":"M. W. Tate, et al., J. Appl. Crystallogr. 28, 196 (1995); Thiel D.J., et al., Rev. Sci. Instrum. 67, 1 (1996).","journal-title":"Rev. Sci. Instrum."},{"key":"e_1_3_1_27_2","unstructured":"Z. Otwinowski in Data Collection and Processing L. Sawyer and S. Bailey Eds. (Science and Engineering Research Council Daresbury Laboratory Daresbury UK 1993) pp. 56\u201362."},{"key":"e_1_3_1_28_2","doi-asserted-by":"crossref","unstructured":"Collaborative Computational Project 4 (CCP4) Acta Crystallogr. D50 760 (1994).","DOI":"10.1107\/S0907444994003112"},{"key":"e_1_3_1_29_2","first-page":"467","volume":"46","author":"Sheldrick G. M.","year":"1990","unstructured":"Sheldrick G. M., ibid46, 467 (1990).","journal-title":"ibid"},{"key":"e_1_3_1_30_2","doi-asserted-by":"crossref","unstructured":"T. A. Jones J. Y. Zou J. Y. Cowan M. Kjeldgaard ibid. A47 110 (1991).","DOI":"10.1107\/S0108767390010224"},{"key":"e_1_3_1_31_2","unstructured":"A. T. Brunger X-PLOR (Version 3.851) Manual (The Howard Hughes Medical Institute and Department of Molecular Biophysics and Biochemistry Yale University New Haven CT)."},{"key":"e_1_3_1_32_2","unstructured":"S. J. Gamblin D. W. Rodgers T. Stehle Proceedings of the CCP4 study weekend Daresbury Laboratory (1996) pp. 163\u2013169."},{"key":"e_1_3_1_33_2","doi-asserted-by":"crossref","unstructured":"K. Y. J. Zhang and P. Main Acta Crystallogr. A46 377.","DOI":"10.1107\/S0108767389012158"},{"key":"e_1_3_1_34_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889891004399"},{"key":"e_1_3_1_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0263-7855(97)00009-X"},{"key":"e_1_3_1_36_2","unstructured":"We thank D. Thiel S. Gruner and members of the MacCHESS staff for support and assistance in data collection at A1; J. Kuriyan S. K. Burley S. Harrison P. Kim E. Gouaux and D. Wang for helpful discussions; Y. Jiang for help in data collection; D. Gadsby and J. Kuriyan for comments on the manuscript; and T. Rahman for patience and support. R.M. is forever grateful to T. Wiesel and A. L. MacKinnon for making this project possible. R.M. is an investigator of the Howard Hughes Medical Institute."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.280.5360.69","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,12]],"date-time":"2024-01-12T23:31:11Z","timestamp":1705102271000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.280.5360.69"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,4,3]]},"references-count":35,"journal-issue":{"issue":"5360","published-print":{"date-parts":[[1998,4,3]]}},"alternative-id":["10.1126\/science.280.5360.69"],"URL":"https:\/\/doi.org\/10.1126\/science.280.5360.69","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,4,3]]}}}