DOI: 10.1128/jb.184.23.6490-6499.2002. Substrate Specificity of the RND-Type Multidrug Efflux Pumps AcrB and AcrD of <i>Escherichia coli</i> Is Determined Predominately by Two Large Periplasmic Loops

Substrate Specificity of the RND-Type Multidrug Efflux Pumps AcrB and AcrD of Escherichia coli Is Determined Predominately by Two Large Periplasmic Loops

10.1128/jb.184.23.6490-6499.2002

Crossref journal-article

American Society for Microbiology

Journal of Bacteriology (235)

Abstract

ABSTRACT AcrAB-TolC is a constitutively expressed, tripartite efflux transporter complex that functions as the primary resistance mechanism to lipophilic drugs, dyes, detergents, and bile acids in Escherichia coli . TolC is an outer membrane channel, and AcrA is an elongated lipoprotein that is hypothesized to span the periplasm and coordinate efflux of such substrates by AcrB and TolC. AcrD is an efflux transporter of E. coli that provides resistance to aminoglycosides as well as to a limited range of amphiphilic agents, such as bile acids, novobiocin, and fusidic acid. AcrB and AcrD belong to the resistance nodulation division superfamily and share a similar topology, which includes a pair of large periplasmic loops containing more than 300 amino acid residues each. We used this knowledge to test several plasmid-encoded chimeric constructs of acrD and acrB for substrate specificity in a marR1 Δ acrB Δ acrD host. AcrD chimeras were constructed in which the large, periplasmic loops between transmembrane domains 1 and 2 and 7 and 8 were replaced with the corresponding loops of AcrB. Such constructs provided resistance to AcrB substrates at levels similar to native AcrB. Conversely, AcrB chimeras containing both loops of AcrD conferred resistance only to the typical substrates of AcrD. These results cannot be explained by simply assuming that AcrD, not hitherto known to interact with AcrA, acquired this ability by the introduction of the loop regions of AcrB, because (i) both AcrD and AcrA were found, in this study, to be required for the efflux of amphiphilic substrates, and (ii) chemical cross-linking in intact cells efficiently produced complexes between AcrD and AcrA. Since AcrD can already interact with AcrA, the alterations in substrate range accompanying the exchange of loop regions can only mean that substrate recognition (and presumably binding) is determined largely by the two periplasmic loops.

Bibliography

Elkins, C. A., & Nikaido, H. (2002). Substrate Specificity of the RND-Type Multidrug Efflux Pumps AcrB and AcrD of Escherichia coli Is Determined Predominately by Two Large Periplasmic Loops. Journal of Bacteriology, 184(23), 6490â6498.

Authors 2

Christopher A. Elkins (first)
Hiroshi Nikaido (additional)

References 27 Referenced 244

Bennett, A. D., and W. V. Shaw. 1983. Resistance to fusidic acid in Escherichia coli mediated by the type I variant of chloramphenicol acetyltransferase. A plasmid-encoded mechanism involving antibiotic binding. Biochem. J.215:29-38. (10.1042/bj2150029) / Biochem. J. (1983)
Bryson, V., and W. Szybalzski. 1952. Microbial selection. Science116:45-51. (10.1126/science.116.3003.45) / Science (1952)
10.1073/pnas.120163297
Davidson, A. L., and H. Nikaido. 1991. Purification and characterization of the membrane-associated components of the maltose transport system from Escherichia coli. J. Biol. Chem.266:8946-8951. (10.1016/S0021-9258(18)31535-7) / J. Biol. Chem. (1991)
Geiser, M., R. Cèbe, D. Drewello, and R. Schmitz. 2001. Integration of PCR fragments at any specific site within cloning vectors without the use of restriction enzymes and DNA ligase. BioTechniques31:88-92. (10.2144/01311st05) / BioTechniques (2001)
10.1128/jb.155.2.531-540.1983
10.1128/JB.183.8.2646-2653.2001
Ma, D., D. N. Cook, M. Alberti, N. G. Pon, J. E. Hearst, and H. Nikaido. 1994. Efflux pumps and drug resistance in gram-negative bacteria. Trends Microbiol.2:489-493. (10.1016/0966-842X(94)90654-8) / Trends Microbiol. (1994)
10.1128/jb.175.19.6299-6313.1993
Ma, D., D. N. Cook, M. Alberti, N. G. Pon, H. Nikaido, and J. E. Hearst. 1995. Genes acrA and acrB encode a stress-induced efflux system of Escherichia coli. Mol. Microbiol.16:45-55. (10.1111/j.1365-2958.1995.tb02390.x) / Mol. Microbiol. (1995)
10.1128/jb.90.1.8-14.1965
10.1128/jb.178.20.5853-5859.1996
Nikaido, H. 1998. Antibiotic resistance caused by gram-negative multidrug efflux pumps. Clin. Infect. Dis.27:S32-S41. (10.1086/514920) / Clin. Infect. Dis. (1998)
Nikaido, H. 2001. Preventing drug access to targets: cell surface permeability barriers and active efflux. Semin. Cell Dev. Biol.12:215-233. (10.1006/scdb.2000.0247) / Semin. Cell Dev. Biol. (2001)
10.1128/JB.180.17.4686-4692.1998
Nikaido, H., and H. I. Zgurskaya. 1999. Antibiotic efflux mechanisms. Curr. Opin. Infect. Dis.12:529-536. (10.1097/00001432-199912000-00001) / Curr. Opin. Infect. Dis. (1999)
10.1128/JB.183.20.5803-5812.2001
10.1128/jb.178.1.306-308.1996
10.1128/jb.155.2.937-939.1983
10.1128/JB.182.6.1754-1756.2000
Saier, M. H., Jr., I. T. Paulsen, M. K. Sliwinski, S. S. Pao, R. A. Skurray, and H. Nikaido. 1998. Evolutionary origins of multidrug and drug-specific efflux pumps in bacteria. FASEB J.12:265-274. / FASEB J. (1998)
10.1128/AAC.45.4.1126-1136.2001
10.1128/JB.184.23.6499-6507.2002
Tseng, T.-T., K. S. Gratwick, J. Kollman, D. Park, D. H. Nies, A. Goffeau, and M. H. Saier, Jr. 1999. The RND permease superfamily: an ancient, ubiquitous and diverse family that includes human disease and development proteins. J. Mol. Microbiol. Biotechnol.1:107-125. / J. Mol. Microbiol. Biotechnol. (1999)
Zgurskaya, H. I., and H. Nikaido. 1999. AcrA from Escherichia coli is a highly asymmetric protein capable of spanning the periplasm. J. Mol. Biol.285:409-420. (10.1006/jmbi.1998.2313) / J. Mol. Biol. (1999)
Zgurskaya, H. I., and H. Nikaido. 1999. Bypassing the periplasm: reconstitution of the AcrAB multidrug efflux pump of Escherichia coli. Proc. Natl. Acad. Sci. USA96:7190-7195. (10.1073/pnas.96.13.7190) / Proc. Natl. Acad. Sci. USA (1999)
10.1128/JB.182.15.4264-4267.2000

Dates

Type	When
Created	22 years, 9 months ago (Nov. 8, 2002, 12:25 p.m.)
Deposited	4 years, 1 month ago (July 29, 2021, 2:06 p.m.)
Indexed	2 days, 20 hours ago (Sept. 3, 2025, 6:05 a.m.)
Issued	22 years, 9 months ago (Dec. 1, 2002)
Published	22 years, 9 months ago (Dec. 1, 2002)
Published Print	22 years, 9 months ago (Dec. 1, 2002)

Funders 0

None

BibTeX

@article{Elkins_2002, title={Substrate Specificity of the RND-Type Multidrug Efflux Pumps AcrB and AcrD of Escherichia coli Is Determined Predominately by Two Large Periplasmic Loops}, volume={184}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.184.23.6490-6499.2002}, DOI={10.1128/jb.184.23.6490-6499.2002}, number={23}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Elkins, Christopher A. and Nikaido, Hiroshi}, year={2002}, month=dec, pages={6490–6498} }

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