TY - JOUR
T1 - An antibiotic-resistance conferring mutation in a neisserial porin
T2 - Structure, ion flux, and ampicillin binding
AU - Bartsch, Annika
AU - Ives, Callum
AU - Kattner, Christof
AU - Pein, Florian
AU - Diehn, Manuel
AU - Tanabe, Mikio
AU - Munk, Axel
AU - Zachariae, Ulrich
AU - Steinem, Claudia
AU - Llabres, Salome
N1 - We are grateful to N. Denkert and M. Meinecke for support in constructing and performing the measurements on black lipid membranes and I. Mey for helpful discussions. Funded by the Deutsche Forschungsgemeinschaft (DFG, Research Foundation Germany)“ under Germany’s Excellence Strategy – EXC 2067/1- 390729940 (C.S. and A.M.), the Bundesministerium für Bildung und Forschung (BMBF) program ZIK HALOmem (FKZ 03Z2HN21 to M.T.), by Grants-in-Aids for young scientist from the MEXT (No. 16K18506 to M.T.), by the Wellcome Trust Interdisciplinary Research Funds (grant WT097818MF), the Scottish Universities’ Physics Alliance (SUPA) and the Tayside Charitable Trust (U.Z. and S.L.), and by the MRC Doctoral Training Programme (C.M.I). The coordinates and the structure factors for the PorB mutant G103K have been deposited in the RCSB Protein Data Bank under the accession code 7DE8.
PY - 2021/6/1
Y1 - 2021/6/1
N2 - Gram-negative bacteria cause the majority of highly drug-resistant bacterial infections. To cross the outer membrane of the complex Gram-negative cell envelope, antibiotics permeate through porins, trimeric channel proteins that enable the exchange of small polar molecules. Mutations in porins contribute to the development of drug-resistant phenotypes. In this work, we show that a single point mutation in the porin PorB from Neisseria meningitidis, the causative agent of bacterial meningitis, can strongly affect the binding and permeation of beta-lactam antibiotics. Using X-ray crystallography, high-resolution electrophysiology, atomistic biomolecular simulation, and liposome swelling experiments, we demonstrate differences in drug binding affinity, ion selectivity and drug permeability of PorB. Our work further reveals distinct interactions between the transversal electric field in the porin eyelet and the zwitterionic drugs, which manifest themselves under applied electric fields in electrophysiology and are altered by the mutation. These observations may apply more broadly to drug-porin interactions in other channels. Our results improve the molecular understanding of porin-based drug-resistance in Gram-negative bacteria.
AB - Gram-negative bacteria cause the majority of highly drug-resistant bacterial infections. To cross the outer membrane of the complex Gram-negative cell envelope, antibiotics permeate through porins, trimeric channel proteins that enable the exchange of small polar molecules. Mutations in porins contribute to the development of drug-resistant phenotypes. In this work, we show that a single point mutation in the porin PorB from Neisseria meningitidis, the causative agent of bacterial meningitis, can strongly affect the binding and permeation of beta-lactam antibiotics. Using X-ray crystallography, high-resolution electrophysiology, atomistic biomolecular simulation, and liposome swelling experiments, we demonstrate differences in drug binding affinity, ion selectivity and drug permeability of PorB. Our work further reveals distinct interactions between the transversal electric field in the porin eyelet and the zwitterionic drugs, which manifest themselves under applied electric fields in electrophysiology and are altered by the mutation. These observations may apply more broadly to drug-porin interactions in other channels. Our results improve the molecular understanding of porin-based drug-resistance in Gram-negative bacteria.
KW - Antibiotic resistance
KW - Beta-barrels
KW - Drug-resistant infections
KW - Electrophysiology
KW - Meningitis
KW - X-ray crystallography
UR - http://www.scopus.com/inward/record.url?scp=85102860078&partnerID=8YFLogxK
U2 - 10.1016/j.bbamem.2021.183601
DO - 10.1016/j.bbamem.2021.183601
M3 - Article
C2 - 33675718
VL - 1863
JO - BBA - Biomembranes
JF - BBA - Biomembranes
SN - 0005-2736
IS - 6
M1 - 183601
ER -