Oligomeric behavior of the RND transporters CusA and AcrB in micellar solution of detergent

被引：13

作者：

Stroebel, David

Sendra, Veronique

Cannella, Dominique

Helbig, Kerstin

Nies, Dietrich H.

Coves, Jacques

机构：

[1] CNRS CEA UJF, UMR 5075, Inst Biol Struct, Lab Prot Membranaires, F-38027 Grenoble, France

[2] Inst Microbiol, D-06099 Halle, Germany

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 2007年 / 1768卷 / 06期

关键词：

RND transporter; analytical ultracentrifugation; detergent; crystallization; membrane protein; oligomer;

D O I：

10.1016/j.bbamem.2007.03.008

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have used analytical ultracentrifugation to explore the oligomeric states of AcrB and CusA in micellar solution of detergent. These two proteins belong to the resistance, nodulation and cell division (RND) family of efflux proteins that are involved in multiple drug and heavy metal resistance. Only the structure of AcrB has been determined so far. Although functional RND proteins should assemble as trimers as AcrB does, both AcrB and CusA form a mixture of quaternary structures (from monomer to heavy oligomer) in detergent solution. The distribution of the oligomeric states was studied as a function of different parameters: nature and concentration of the detergent, ionic strength, pH, protein concentration. This pseudo-heterogeneity does not hamper the crystallization of AcrB as a homotrimer. (c) 2007 Elsevier B.V. All rights reserved.

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页码：1567 / 1573

页数：7

相关论文

共 29 条

[1] Macromolecular size-and-shape distributions by sedimentation velocity analytical ultracentrifugation [J].

Brown, Patrick H. ;

Schuck, Peter .

BIOPHYSICAL JOURNAL, 2006, 90 (12) :4651-4661

[2] Purification of Na+,K+-ATPase expressed in Pichia pastoris reveals an essential role of phospholipid-protein interactions [J].

Cohen, E ;

Goldshleger, R ;

Shainskaya, A ;

Tal, DM ;

Ebel, C ;

le Maire, M ;

Karlish, SJD .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (17) :16610-16618

[3] A rapid method for assessing lipid:protein and detergent:protein ratios in membrane-protein crystallization [J].

daCosta, CJB ;

Baenziger, JE .

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 2003, 59 :77-83

[4] Calculation of hydrodynamic properties of globular proteins from their atomic-level structure [J].

de la Torre, JG ;

Huertas, ML ;

Carrasco, B .

BIOPHYSICAL JOURNAL, 2000, 78 (02) :719-730

[5] Effect of detergents on the association of the glycophorin A transmembrane helix [J].

Fisher, LE ;

Engelman, DM ;

Sturgis, JN .

BIOPHYSICAL JOURNAL, 2003, 85 (05) :3097-3105

[6] POLYRIBOSOME ANALYSIS ON SUCROSE GRADIENTS PRODUCED BY THE FREEZE-THAW METHOD [J].

FOURCROY, P ;

CUILLER, S ;

LARGITTE, FC ;

LAMBERT, C .

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 1981, 4 (3-4) :243-246

[7] Molecular analysis of the copper-transporting efflux system CusCFBA of Escherichia coli [J].

Franke, S ;

Grass, G ;

Rensing, C ;

Nies, DH .

JOURNAL OF BACTERIOLOGY, 2003, 185 (13) :3804-3812

[8] Energetics and topology of CzcA, a cation/proton antiporter of the resistance-nodulation-cell division protein family [J].

Goldberg, M ;

Pribyl, T ;

Juhnke, S ;

Nies, DH .

JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (37) :26065-26070

[9] The BPTI decamer observed in acidic pH crystal forms pre-exists as a stable species in solution [J].

Hamiaux, C ;

Pérez, J ;

Prangé, T ;

Veesler, S ;

Riès-Kautt, M ;

Vachette, P .

JOURNAL OF MOLECULAR BIOLOGY, 2000, 297 (03) :697-712

[10] Oligomeric states of the detergent-solubilized human serum paraoxonase (PON1) [J].

Josse, D ;

Ebel, C ;

Stroebel, D ;

Fontaine, A ;

Borges, F ;

Echalier, A ;

Baud, D ;

Renault, F ;

le Maire, M ;

Chabrières, E ;

Masson, P .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (36) :33386-33397