Deleting two C-terminal α-helices is effective to crystallize the bacterial ABC transporter Escherichia coli MsbA complexed with AMP-PNP

被引：5

作者：

Terakado, Kanako ^{[1
,2
]}

Kodan, Atsushi ^{[2
,3
]}

Nakano, Hiroaki ^{[1
]}

Kimura, Yasuhisa ^{[2
,4
]}

Ueda, Kazumitsu ^{[3
,4
]}

Nakatsu, Toru ^{[1
,2
]}

Kato, Hiroaki ^{[1
,2
]}

机构：

[1] Kyoto Univ, Grad Sch Pharmaceut Sci, Dept Biol Struct, Kyoto 6068501, Japan

[2] RIKEN Harima Inst SPring 8, Sayo, Hyogo 6795148, Japan

[3] Kyoto Univ, Inst Integrated Cell Mat Sci, Kyoto 6068502, Japan

[4] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Lab Cellular Biochem, Kyoto 6068502, Japan

来源：

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2010年 / 66卷

关键词：

P-GLYCOPROTEIN; INNER MEMBRANE; GENE; TRANSLOCATION; RESISTANCE; SURFACE; FAMILY; DOMAIN;

D O I：

10.1107/S0907444909055504

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：319 / 323

页数：5