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ヨコヤマ ケン
YOKOYAMA KEN
横山 謙 所属 京都産業大学 生命科学部 先端生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2006/02 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | Two-dimensional crystallization and analysis of projection images of intact Thermus thermophilus V-ATPase |
| 執筆形態 | その他 |
| 掲載誌名 | JOURNAL OF STRUCTURAL BIOLOGY |
| 出版社・発行元 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| 巻・号・頁 | 153(2),pp.200-206 |
| 著者・共著者 | C Gerle,K Tani,K Yokoyama,M Tamakoshi,M Yoshida,Y Fujiyoshi,K Mitsuoka |
| 概要 | H+-ATPase/synthases are membrane-bound rotary nanomotors that are essential for energy conversion in nearly all life forms. A member of the family of the vacuolar-type ATPases (V-ATPases) from Thermus thermophilus, sometimes also termed A-type ATPase, was purified to homogeneity and subjected to two-dimensional (2D) crystallization trials, A novel approach to the 2D crystallization of unstable complexes yielded densely packed sheets or V-ATPase, exhibiting crystalline arrays, Aggregation of the V-ATPase under acidic conditions during reconstitution circumvented the continuous dissociation of the whole complex into the I-1' and I-o' domains. The resulting three-dimensional aggregates were converted into 2D sheets by the use ora basic buffer, and after it short annealing cycle, ordered arrays of up to 1.5 mu m diameter appeared. Fourier transforms calculated from micrographs taken from the negatively stained sample showed diffraction spots to a resolution of 23 angstrom. The Fourier transforms of the untilted images revealed unit-cell dimensions of a = 232 angstrom, b = 132 angstrom, and gamma = 90 degrees, and a projection map was calculated by merging 111 images. The most probable molecular packing suggests p22(1)2(1) symmetry of the crystals and dimer contacts between the V-1 domains. (C) 2005 Elsevier Inc. All rights reserved. |
| DOI | 10.1016/j.jsb.2005.11.004 |
| ISSN | 1047-8477/1095-8657 |
| PMID | 16377206 |