ジャーナル: Nature / 年: 2014 タイトル: Structure of the AcrAB-TolC multidrug efflux pump. 著者: Dijun Du / Zhao Wang / Nathan R James / Jarrod E Voss / Ewa Klimont / Thelma Ohene-Agyei / Henrietta Venter / Wah Chiu / Ben F Luisi / 要旨: The capacity of numerous bacterial species to tolerate antibiotics and other toxic compounds arises in part from the activity of energy-dependent transporters. In Gram-negative bacteria, many of ...The capacity of numerous bacterial species to tolerate antibiotics and other toxic compounds arises in part from the activity of energy-dependent transporters. In Gram-negative bacteria, many of these transporters form multicomponent 'pumps' that span both inner and outer membranes and are driven energetically by a primary or secondary transporter component. A model system for such a pump is the acridine resistance complex of Escherichia coli. This pump assembly comprises the outer-membrane channel TolC, the secondary transporter AcrB located in the inner membrane, and the periplasmic AcrA, which bridges these two integral membrane proteins. The AcrAB-TolC efflux pump is able to transport vectorially a diverse array of compounds with little chemical similarity, thus conferring resistance to a broad spectrum of antibiotics. Homologous complexes are found in many Gram-negative species, including in animal and plant pathogens. Crystal structures are available for the individual components of the pump and have provided insights into substrate recognition, energy coupling and the transduction of conformational changes associated with the transport process. However, how the subunits are organized in the pump, their stoichiometry and the details of their interactions are not known. Here we present the pseudo-atomic structure of a complete multidrug efflux pump in complex with a modulatory protein partner from E. coli. The model defines the quaternary organization of the pump, identifies key domain interactions, and suggests a cooperative process for channel assembly and opening. These findings illuminate the basis for drug resistance in numerous pathogenic bacterial species.
履歴
登録
2013年10月31日
登録サイト: PDBE / 処理サイト: PDBE
改定 1.0
2014年4月30日
Provider: repository / タイプ: Initial release
改定 1.1
2014年5月28日
Group: Database references
改定 1.2
2015年9月23日
Group: Data collection
改定 1.3
2019年5月8日
Group: Data collection / Experimental preparation / Other カテゴリ: exptl_crystal_grow / pdbx_database_proc / pdbx_database_status Item: _exptl_crystal_grow.temp / _pdbx_database_status.recvd_author_approval
温度: 293 K / 手法: 蒸気拡散法, ハンギングドロップ法 / pH: 7.4 詳細: THE ACRBZ COMPLEX AT 10 MG ML-1 USING SAMPLE BUFFER. 9 MM N-OCTYL-BETA-D-THIOGLUCOPYRANOSIDE (90 MM) WAS MIXED WITH ACRBZ COMPLEX BEFORE THE CRYSTALLISATION TRIALS. THE ACRBZ CRYSTALS WERE ...詳細: THE ACRBZ COMPLEX AT 10 MG ML-1 USING SAMPLE BUFFER. 9 MM N-OCTYL-BETA-D-THIOGLUCOPYRANOSIDE (90 MM) WAS MIXED WITH ACRBZ COMPLEX BEFORE THE CRYSTALLISATION TRIALS. THE ACRBZ CRYSTALS WERE GROWN AT 20 C USING THE HANGING-DROPLET VAPOUR DIFFUSION METHOD BY MIXING 4 MICROLITERS OF ACRBZ COMPLEX WITH 2 MICROLITERS OF RESERVOIR SOLUTION (100 MM TRICINE PH: 7.4, 50 MM LITHIUM SULPHATE, 5 MM CADMIUM CHLORIDE HYDRATE, 7 % PEG 3000, 10% GLYCEROL).
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データ収集
回折
ID
平均測定温度 (K)
Crystal-ID
1
100
1
2
100
1
放射光源
由来
サイト
ビームライン
ID
波長
シンクロトロン
Diamond
I24
1
0.9794
シンクロトロン
Diamond
I04-1
2
0.9794
検出器
タイプ
ID
検出器
日付
DECTRIS PILATUS 2M
1
PIXEL
2013年1月10日
DECTRIS PIXEL
2
PIXEL
2013年1月10日
放射
ID
プロトコル
単色(M)・ラウエ(L)
散乱光タイプ
Wavelength-ID
1
SINGLEWAVELENGTH
M
x-ray
1
2
SINGLEWAVELENGTH
M
x-ray
1
放射波長
波長: 0.9794 Å / 相対比: 1
反射
解像度: 3.69→24.89 Å / Num. obs: 23077 / % possible obs: 94.5 % / Observed criterion σ(I): 2 / 冗長度: 10.6 % / Biso Wilson estimate: 148 Å2 / Rmerge(I) obs: 0.13 / Net I/σ(I): 15.1
反射 シェル
解像度: 3.69→3.99 Å / 冗長度: 10.6 % / Rmerge(I) obs: 1.17 / Mean I/σ(I) obs: 2.1 / % possible all: 94.8
解像度: 3.7→24.91 Å / Cor.coef. Fo:Fc: 0.864 / Cor.coef. Fo:Fc free: 0.858 / SU B: 54.668 / SU ML: 0.78 / 交差検証法: THROUGHOUT / ESU R Free: 0.844 / 立体化学のターゲット値: MAXIMUM LIKELIHOOD / 詳細: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS.
Rfactor
反射数
%反射
Selection details
Rfree
0.36127
1161
5 %
RANDOM
Rwork
0.33906
-
-
-
obs
0.34009
21912
94.04 %
-
溶媒の処理
イオンプローブ半径: 0.8 Å / 減衰半径: 0.8 Å / VDWプローブ半径: 1.2 Å / 溶媒モデル: MASK