Structure of the cytoplasmic β subunit - T1 assembly of voltage- dependent K+ channels

Science

Volumn 289, Issue 5476, 2000, Pages 123-127

  Gulbis, Jacqueline M ^a,b   Zhou, Ming ^a   Mann, Sabine ^a   MacKinnon, Roderick ^a  

^a ROCKEFELLER UNIVERSITY   (United States)

^b CSIRO   (Australia)

Author keywords

[No Author keywords available]

Indexed keywords

POTASSIUM CHANNEL; PROTEIN SUBUNIT;

ARTICLE; HUMAN; NONHUMAN; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN ASSEMBLY; PROTEIN STRUCTURE; STRUCTURE ANALYSIS; X RAY CRYSTALLOGRAPHY;

ANIMALS; CELL LINE; CRYSTALLOGRAPHY, X-RAY; CYTOPLASM; KV1.1 POTASSIUM CHANNEL; KV1.4 POTASSIUM CHANNEL; MACROMOLECULAR SUBSTANCES; MODELS, MOLECULAR; MUTATION; OOCYTES; OXIDOREDUCTASES; PATCH-CLAMP TECHNIQUES; PEPTIDES; POTASSIUM CHANNELS; POTASSIUM CHANNELS, VOLTAGE-GATED; PROTEIN CONFORMATION; PROTEIN STRUCTURE, QUATERNARY; PROTEIN STRUCTURE, TERTIARY; RATS; RECOMBINANT FUSION PROTEINS; XENOPUS;

EID: 0034617121     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.289.5476.123     Document Type: Article

References (34)

1. J. M. Gulbis, S. Mann, R. MacKinnon, Cell 97, 943 (1999).
2. G. Shi et al., Neuron 16, 843 (1996).
3. N. Nagaya and D. M. Papazian, J. Biol. Chem. 272, 3022 (1997).
4. A. Kreusch, P. J. Pfaffinger, C. F. Stevens, S. Choe, Nature 392, 945 (1998).
5. C. M. Armstrong, J. Gen. Physiol. 58, 413 (1971).
6. W. N. Zagotta, T. Hoshi, R. W. Aldrich, Science 250, 568 (1990).
7. S. D. Demo and C. Yellen, Neuron 7, 743 (1991).
8. S. Sewing, J. Roeper, O. Pongs, Neuron 16, 455 (1996).
9. W. Yu, J. Xu, M. Li, Neuron 16, 441 (1996).
10. J. Rettig et al., Nature 369, 289 (1994).
11. A. Baumann, A. Grupe, A. Ackermann, O. Pongs, EMBO J. 7, 2457 (1988).
12. free was monitored to reduce model bias. Individual temperature factors were refined for all nonhydrogen atoms.
13. free was monitored to reduce model bias. Individual temperature factors were refined for all nonhydrogen atoms.
14. free was monitored to reduce model bias. Individual temperature factors were refined for all nonhydrogen atoms.
15. free was monitored to reduce model bias. Individual temperature factors were refined for all nonhydrogen atoms.
16. 2, and 5 mM Hepes (pH 7.6) with NaOH. Analog data from the amplifier were filtered (2 kHz, - 3 dB) with an eight-pole Bessel filter (Frequency Devices, Haverhill, MA), digitized at 10 kHz, and transferred to a computer hard disk with an Axon 1200 digitization board and supporting software (Clampex version 8; Axon Instrument, Foster City, CA).
17. T. E. Lee, L. H. Philipson, D. J. Nelson, J. Membr. Biol. 151, 225 (1996).
18. J. M. Gulbis, M. Zhou, S. Mann, R. MacKinnon, data not shown.
19. K. Nakahira, G. Shi, K. J. Rhodes, J. S. Trimmer, J. Biol. Chem. 271, 7084 (1996).
20. R. D. Murrell-Langnado and R. W. Aldrich, J. Gen. Physiol. 102, 949 (1993).
21. Inactivation time constants were obtained by fitting the current decay with exponential functions with Clampfit (Axon) software. Expression of Kv1.4-IR with β12 chimera yielded two exponentials: a predominant (>90%) fast component (time τ < 60 ms) and a minor (< 10%) component (τ ∼ 500 ms). The slower time constant was relatively invariable and coincided with the COOH-type inactivation of Kv1.4-IR [S. H. Heinemann, J. Rettig, H. R. Graack, O. Pongs, J. Physiol. 493, 625 (1996)]. In double-mutant cycle analysis, only the predominant component was used. Inactivation recovery was studied with paired pulses (22). The peak of the second pulse in the pair was graphed as a function of time after the first pulse, defining an envelope of recovery that was fit to an exponential function with Origin (Microcol Software, Northampton, MA). Recovery from inactivation for Kv1.4-IR paired with the β12 chimera was well described by single exponential function.
22. d is very close to the equilibrium ratio of open to inactivated channels. A value of Ω greater than unity indicates that the effects of two mutations are coupled.
23. W. R. Kobertz and C. Miller, Nature Struct. Biol. 6, 1122 (1999).
24. A. Miyazawa, Y. Fujiyoshi, M. Stowell, N. Unwin, J. Mol. Biol. 288, 765 (1999).
25. R. MacKinnon, R. W. Aldrich, A. W. Lee, Science 262, 757 (1993).
26. F. Gomez-Lagunas and C. M. Armstrong, Biophys. J. 68, 89 (1995).
27. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
28. T. A. Jones, J. Y. Zou, S. W. Cowan, M. Kjeldgaard, Acta Crystallogr. Sect. A 47, 110 (1991).
29. P. Kraulis, J. Appl. Crystallogr. 24, 946 (1991).
30. D. Bacon and W. F. Anderson, J. Mol. Graph 6, 219 (1988).
31. See www.povray.org.
32. A. Nicholls, K. A. Sharp, B. Honig, Proteins 11, 281 (1991).
33. D. A. Doyle et al., Science 280, 69 (1998).
34. We thank J. Bonnano for technical assistance; J. Morais Cabral, R. Dutzler, Y. Jiang, A. Pico, and Y. Zhou for help with data collection; B. Chait and M. Cadene for mass spectrometry; O. Pongs for Kv1.4 DNA; J. Trimmer for β subunit DNA; members of the MacCHESS (A1) and Brookhaven National Laboratory (X25) staff for assistance in data collection; and W. Chin for artwork and help with the manuscript. The coordinates have been submitted under Protein Data Bank ID code 1EXB. R.M. is an investigator in the Howard Hughes Medical Institute. Supported by NIH grant GM47400.