| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Ornik, Alina R. Sjuts, Hanno Nguyen, Son T. Kim, Hong-suk Opperman, Timothy J. Ding, Xiaoyuan Vargiu, Attilio V. Kwasny, Steven M. Pos, Klaas M. Nikaido, Hiroshi Ruggerone, Paolo Bowlin, Terry L. |
| Description |
Author Affiliation: Sjuts H ( Institute of Biochemistry, Goethe University Frankfurt, D-60438 Frankfurt, Germany); Vargiu AV ( Department of Physics, University of Cagliari, I-09042 Monserrato (CA), Italy); Kwasny SM ( Department of Molecular and Cell Biology, Microbiotix, Inc., Worcester, MA 01605); Nguyen ST ( Department of Molecular and Cell Biology, Microbiotix, Inc., Worcester, MA 01605); Kim HS ( University of California, Berkeley, CA 94720-3202.); Ding X ( Department of Molecular and Cell Biology, Microbiotix, Inc., Worcester, MA 01605); Ornik AR ( Institute of Biochemistry, Goethe University Frankfurt, D-60438 Frankfurt, Germany); Ruggerone P ( Department of Physics, University of Cagliari, I-09042 Monserrato (CA), Italy); Bowlin TL ( Department of Molecular and Cell Biology, Microbiotix, Inc., Worcester, MA 01605); Nikaido H ( University of California, Berkeley, CA 94720-3202 vargiu@dsf.unica.it nhiroshi@berkeley.edu pos@em.uni-frankfurt.de topperman@microbiotix.com.); Pos KM ( Institute of Biochemistry, Goethe University Frankfurt, D-60438 Frankfurt, Germany); Opperman TJ ( Department of Molecular and Cell Biology, Microbiotix, Inc., Worcester, MA 01605); |
| Abstract |
The Escherichia coli AcrAB-TolC efflux pump is the archetype of the resistance nodulation cell division (RND) exporters from Gram-negative bacteria. Overexpression of RND-type efflux pumps is a major factor in multidrug resistance (MDR), which makes these pumps important antibacterial drug discovery targets. We have recently developed novel pyranopyridine-based inhibitors of AcrB, which are orders of magnitude more powerful than the previously known inhibitors. However, further development of such inhibitors has been hindered by the lack of structural information for rational drug design. Although only the soluble, periplasmic part of AcrB binds and exports the ligands, the presence of the membrane-embedded domain in AcrB and its polyspecific binding behavior have made cocrystallization with drugs challenging. To overcome this obstacle, we have engineered and produced a soluble version of AcrB [AcrB periplasmic domain (AcrBper)], which is highly congruent in structure with the periplasmic part of the full-length protein, and is capable of binding substrates and potent inhibitors. Here, we describe the molecular basis for pyranopyridine-based inhibition of AcrB using a combination of cellular, X-ray crystallographic, and molecular dynamics (MD) simulations studies. The pyranopyridines bind within a phenylalanine-rich cage that branches from the deep binding pocket of AcrB, where they form extensive hydrophobic interactions. Moreover, the increasing potency of improved inhibitors correlates with the formation of a delicate protein- and water-mediated hydrogen bond network. These detailed insights provide a molecular platform for the development of novel combinational therapies using efflux pump inhibitors for combating multidrug resistant Gram-negative pathogens. |
| ISSN |
00278424 |
| e-ISSN |
10916490 |
| Journal |
Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number |
13 |
| Volume Number |
113 |
| Language |
English |
| Publisher |
National Academy of Sciences |
| Publisher Date |
2016-04-01 |
| Publisher Place |
United States |
| Access Restriction |
Open |
| Subject Keyword |
Anti-Bacterial Agents Pharmacology Escherichia Coli Proteins Antagonists & Inhibitors Multidrug Resistance-Associated Proteins Pyridines Chemistry Binding Sites Crystallography, X-Ray Drug Discovery Drug Resistance, Multiple, Bacterial Drug Effects Escherichia Coli Metabolism Hydrophobic And Hydrophilic Interactions Models, Molecular Molecular Dynamics Simulation Protein Structure, Tertiary Pyrans Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type |
Text |
| Resource Type |
Article |
| Subject |
Multidisciplinary |
