2010年3月
Deleting two C-terminal alpha-helices is effective to crystallize the bacterial ABC transporter Escherichia coli MsbA complexed with AMP-PNP
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
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- 巻
- 66
- 号
- 3
- 開始ページ
- 319
- 終了ページ
- 323
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1107/S0907444909055504
- 出版者・発行元
- WILEY-BLACKWELL PUBLISHING, INC
An MsbA deletion mutant Delta C21 that lacks the two C-terminal alpha-helices was expressed in Escherichia coli strain C41 and purified by metal-affinity and gel-filtration chromatography. Purified Delta C21 retained 26% of the activity of the wild-type ATPase and had a similar binding affinity to fluorescent nucleotide derivatives. Although crystals of wild-type MsbA complexed with adenosine 5'-(beta,gamma-imido) triphosphate could not be obtained, crystals of Delta C21 that diffracted to 4.5 angstrom resolution were obtained. The preliminary Delta C21 structure had the outward-facing conformation, in contrast to the previously reported E. coli MsbA structure. This result suggests that deletion of the C-terminal alpha-helices may play a role in facilitating the outward-facing nucleotide-bound crystal structure of EcMsbA.
- リンク情報
- ID情報
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- DOI : 10.1107/S0907444909055504
- ISSN : 0907-4449
- PubMed ID : 20179345
- Web of Science ID : WOS:000274536400013