In vitro transport activity of the fully assembled MexAB-OprM efflux pump from Pseudomonas aeruginosa

Nature Communications

Volumn 6, Issue , 2015, Pages

  Verchère, Alice ^a   Dezi, Manuela ^a,c   Adrien, Vladimir ^b   Broutin, Isabelle ^a   Picard, Martin ^a  

^a UNIVERSITÉ PARIS DESCARTES   (France)

^b SORBONNE UNIVERSITÉ   (France)

^c UNIVERSITÉ PIERRE ET MARIE CURIE   (France)

Author keywords

[No Author keywords available]

Indexed keywords

ANTIBIOTIC AGENT; BACTERIAL PROTEIN; ETHIDIUM BROMIDE; HYDROCHLORIC ACID; LIPOSOME; MEXA PROTEIN; MEXB PROTEIN; OPRM PROTEIN; PROTEOLIPOSOME; UNCLASSIFIED DRUG; ANTIINFECTIVE AGENT; CARRIER PROTEIN; ETHIDIUM; FLUORESCENT DYE; MEROPENEM; MEXA PROTEIN, PSEUDOMONAS AERUGINOSA; MEXB PROTEIN, PSEUDOMONAS AERUGINOSA; OPRM PROTEIN, PSEUDOMONAS AERUGINOSA; OUTER MEMBRANE PROTEIN; PROTEOLIPID; PROTEOLIPOSOMES; THIENAMYCIN DERIVATIVE;

ANTIBIOTIC RESISTANCE; BIOASSAY; MEMBRANE; MICROBIAL ACTIVITY; PUBLIC HEALTH;

ARTICLE; CYTOPLASM; FLUORESCENCE; IN VITRO STUDY; NONHUMAN; PH; PROTEIN TRANSPORT; PSEUDOMONAS AERUGINOSA; ANTIBIOTIC RESISTANCE; METABOLISM;

NEGIBACTERIA; PSEUDOMONAS AERUGINOSA;

ANTI-BACTERIAL AGENTS; BACTERIAL OUTER MEMBRANE PROTEINS; DRUG RESISTANCE, BACTERIAL; ETHIDIUM; FLUORESCENT DYES; HYDROGEN-ION CONCENTRATION; IN VITRO TECHNIQUES; MEMBRANE TRANSPORT PROTEINS; PROTEOLIPIDS; PSEUDOMONAS AERUGINOSA; THIENAMYCINS;

EID: 84928781008     PISSN: None     EISSN: 20411723     Source Type: Journal    
DOI: 10.1038/ncomms7890     Document Type: Article

References (27)

1. May, M. Drug development: time for teamwork. Nature 509, S4-S5 (2014).
2. Stover, C. K. et al. Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406, 1-6 (2000).
3. Sennhauser, G., Bukowska, M. A., Briand, C. & Grutter, M. G. Crystal structure of the multidrug exporter MexB from Pseudomonas aeruginosa. J. Mol. Biol. 389, 134-145 (2009).
4. Akama, H. et al. Crystal structure of the membrane fusion protein, MexA, of the multidrug transporter in Pseudomonas aeruginosa. J. Biol. Chem. 279, 25939-25942 (2004).
5. Symmons, M. F., Bokma, E., Koronakis, E., Hughes, C. & Koronakis, V. The assembled structure of a complete tripartite bacterial multidrug efflux pump. Proc. Natl Acad. Sci. 106, 7173-7178 (2009).
6. Phan, G. et al. Structural and dynamical insight into the opening mechanism of P. aeruginosa OprM channel. Structure 18, 507-517 (2010).
7. Lomovskaya, O., Zgurskaya, H. I., Totrov, M. & Watkins, W. J. Waltzing transporters and 'the dance macabre' between humans and bacteria. Nat. Rev. Drug Discov. 6, 56-65 (2006).
8. Markham, P. & Neyfakh, N. Inhibition of the multidrug transporter NorA prevents emergence of norfloxacin resistance in Staphylococcus aureus. Antimicrob. Agents. Chemother. 40, 2673-2674 (1996).
9. Zgurskaya, H. I. & Nikaido, H. Bypassing the periplasm: reconstitution of the AcrAB multidrug efflux pump of Escherichia coli. Proc. Natl Acad. Sci. USA 96, 7190-7195 (1999).
10. Goldberg, M., Pribyl, T. & Juhnke, S. Energetics and topology of CzcA, a cation/proton antiporter of the resistance-nodulation-cell division protein family. J. Biol. Chem. 274, 26065-26070 (1999).
11. Long, F. et al. Crystal structures of the CusA efflux pump suggest methioninemediated metal transport. Nature 467, 484-488 (2010).
12. Welch, A., Awah, C. U., Jing, S., van Veen, H. W. & Venter, H. Promiscuous partnering and independent activity of MexB, the multidrug transporter protein from Pseudomonas aeruginosa. Biochem. J. 430, 355-364 (2010).
13. Verchère, A., Broutin, I. & Picard, M. Hoechst likes to play hide and seek: use it with caution! Anal. Biochem. 440, 117-119 (2013).
14. Aires, J. R. & Nikaido, H. Aminoglycosides are captured from both periplasm and cytoplasm by the AcrD multidrug efflux transporter of Escherichia coli. J. Bacteriol. 187, 1923-1929 (2005).
15. Picard, M., Verchère, A. & Broutin, I. Monitoring the active transport of efflux pumps after their reconstitution into proteoliposomes: caveats and keys. Anal. Biochem. 420, 194-196 (2012).
16. Verchère, A., Broutin, I. & Picard, M. Photo-induced proton gradients for the in vitro investigation of bacterial efflux pumps. Sci. Rep. 2, 306 (2012).
17. Verchère, A., Dezi, M., Broutin, I. & Picard, M. In vitro investigation of the MexAB efflux pump from Pseudomonas aeruginosa. J. Vis. Exp. e50894 (2014).
18. Sedgwick, E. G. & Bragg, P. D. Differential movement of ions in artificial phospholipid vesicles. FEBS Lett. 272, 81-84 (1990).
19. Livermore, D. M. Multiple mechanisms of antimicrobial resistancein Pseudomonas aeruginosa: our worst nightmare? Antimicrob. Res. 34, 895-910 (2002).
20. Trépout, S. et al. Structure of reconstituted bacterial membrane efflux pump by cryo-electron tomography. Biochim. Biophys. Acta 1798, 1953-1960 (2010).
21. Schulz, R., Vargiu, A. V., Collu, F., KleinekathOfer, U. & Ruggerone, P. Functional rotation of the transporter AcrB: insights into drug extrusion from simulations. PLoS Comput. Biol. 6, e1000806 (2010).
22. Imai, T. et al. Functionality mapping on internal surfaces of multidrug transporter AcrB based on molecular theory of solvation: implications for drug efflux pathway. J. Phys. Chem. B 115, 8288-8295 (2011).
23. Pos, K. M. Drug transport mechanism of the AcrB efflux pump. Biochim. Biophys. Acta 1794, 782-793 (2009).
24. Mokhonov, V. et al. Multidrug transporter MexB of Pseudomonas aeruginosa: overexpression, purification, and initial structural characterization. Protein Expr. Purif. 40, 91-100 (2005).
25. Ponchon, L. et al. Co-expression of RNA-protein complexes in Escherichia coli and applications to RNA biology. Nucleic Acids Res. 41, e150 (2013).
26. Wang, R. E., ZHANG, Y., Cai, J., Cai, W. & Gao, T. Aptamer-based fluorescent biosensors. Curr. Med. Chem. 18, 4175-4184 (2011).
27. Nutiu, R. & Li, Y. Aptamers with fluorescence-signaling properties. Methods 37, 16-25 (2005).