MOMP from Campylobacter jejuni Is a Trimer of 18-Stranded β-Barrel Monomers with a Ca2+Ion Bound at the Constriction Zone.
The Gram-negative organism Campylobacter jejuni is the major cause of food poisoning. Unlike Escherichia coli, which has two major porins, OmpC and OmpF, C. jejuni has one, termed major outer membrane protein (MOMP) through which nutrients and antibiotics transit. We report the 2.1-Å crystal structure of C. jejuni MOMP expressed in E. coli and a lower resolution but otherwise identical structure purified directly from C. jejuni. The 2.1-Å resolution structure of recombinant MOMP showed that although the protein has timeric arrangement similar to OmpC, it is an 18-stranded, not 16-stranded, β-barrel. The structure has identified a Ca2+bound at the constriction zone, which is functionally significant as suggested by molecular dynamics and single-channel experiments. The water-filled channel of MOMP has a narrow constriction zone, and single-molecule studies show a monomeric conductivity of 0.7±0.2 nS and a trimeric conductance of 2.2±0.2 nS. The ion neutralizes negative charges at the constriction zone, reducing the transverse electric field and reversing ion selectivity. Modeling of the transit of ciprofloxacin, an antibiotic of choice for treating Campylobacter infection, through the pore of MOMP reveals a trajectory that is dependent upon the presence metal ion.
Biomedical Sciences Research Complex, University of St Andrews, 09042 St Andrews, UK.
Luana G M
LGM
Ferrara
Biomedical Sciences Research Complex, University of St Andrews, 09042 St Andrews, UK.
Gregor D
GD
Wallat
Biomedical Sciences Research Complex, University of St Andrews, 09042 St Andrews, UK.
Lucile
L
Moynié
Department of Life Sciences and Chemistry, Jacobs University Bremen, 28719 Bremen, Germany.
Naresh N
NN
Dhanasekar
Aix Marseille Univ, IRBA, TMCD2, 13385 Marseille, France.
Soumeya
S
Aliouane
Department of Physics, University of Cagliari, Cittadella Universitaria Monserrato, S.P8-km 0.700, 09042 Monserrato, Cagliari (CA), Italy.
Silvia
S
Acosta-Gutiérrez
Aix Marseille Univ, IRBA, TMCD2, 13385 Marseille, France.
Jean-Marie
JM
Pagès
Aix Marseille Univ, IRBA, TMCD2, 13385 Marseille, France.
Jean-Michel
JM
Bolla
Department of Life Sciences and Chemistry, Jacobs University Bremen, 28719 Bremen, Germany.
Mathias
M
Winterhalter
Department of Physics, University of Cagliari, Cittadella Universitaria Monserrato, S.P8-km 0.700, 09042 Monserrato, Cagliari (CA), Italy.
Matteo
M
Ceccarelli
Biomedical Sciences Research Complex, University of St Andrews, 09042 St Andrews, UK; State Key Laboratory of Biotherapy, Sichuan University, Chengdu 610041, China. Electronic address: naismith@st-andrews.ac.uk.
James H
JH
Naismith
D000900:Anti-Bacterial Agents / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D016123:Campylobacter jejuni / Q000737:chemistry*; D002939:Ciprofloxacin / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018272:Porins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D046912:Multiprotein Complexes; D018272:Porins; D011994:Recombinant Proteins; C095080:major outer membrane protein, Campylobacter jejuni; D002939:Ciprofloxacin; D002118:Calcium
Campylobacter; antibiotic resistance; outer membrane proteins; porins; β-barrel
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