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From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins
Published on: July 4, 2016
Identification and characterization of a novel outer membrane protein (OMP J) of Moraxella catarrhalis that exists in
John P Hays1, Saskia van Selm, Theo Hoogenboezem
1Department of Medical Microbiology and Infectious Diseases, Erasmus MC, Rotterdam, The Netherlands. j.hays@erasmusmc.nl
Abstract:
Moraxella catarrhalis is a common commensal of the human respiratory tract that has been associated with a number of disease states, including acute otitis media in children and exacerbations of chronic obstructive pulmonary disease in adults. During studies to investigate the outer membrane proteins of this bacterium, two novel major proteins, of approximately 19 kDa and 16 kDa (named OMP J1 and OMP J2, respectively), were identified. Further analysis indicated that these two proteins possessed almost identical gene sequences, apart from two insertion/deletion events in predicted external loops present within the putative barrel-like structure of the proteins. The development of a PCR screening strategy found a 100% (96/96) incidence for the genes encoding the OMP J1 and OMP J2 proteins within a set of geographically diverse M. catarrhalis isolates, as well as a significant association of OMP J1/OMP J2 with both the genetic lineage and the complement resistance phenotype (Fisher's exact test; P < 0.01). Experiments using two DeltaompJ2 mutants (one complement resistant and the other complement sensitive) indicated that both were less easily cleared from the lungs of mice than were their isogenic wild-type counterparts, with a significant difference in bacterial clearance being observed for the complement-resistant isolate but not for its isogenic DeltaompJ2 mutant (unpaired Student's t test; P < 0.001 and P = 0.32). In this publication, we characterize a novel outer membrane protein of Moraxella catarrhalis which exists in two variant forms associated with particular genetic lineages, and both forms are suggested to contribute to bacterial clearance from the lungs.
Insights
Two novel outer membrane proteins, OMP J1 and OMP J2, were identified in Moraxella catarrhalis. These proteins are linked to bacterial lineage and complement resistance, aiding in lung clearance.
Area of Science:
- Microbiology
- Bacterial Pathogenesis
- Immunology
Background:
- Moraxella catarrhalis is a common human respiratory tract commensal.
- It is associated with diseases like otitis media and COPD exacerbations.
- Outer membrane proteins are crucial for bacterial-host interactions.
Purpose of the Study:
- To characterize novel outer membrane proteins (OMPs) of Moraxella catarrhalis.
- To investigate the genetic and functional significance of these OMPs.
- To determine the role of these OMPs in bacterial virulence and host immune evasion.
Main Methods:
- Identification and characterization of novel OMPs (OMP J1 and OMP J2) using protein analysis.
- PCR screening to determine gene incidence in diverse M. catarrhalis isolates.
- Construction and testing of DeltaompJ2 mutants in a murine lung clearance model.
Main Results:
- Two novel OMPs, OMP J1 (19 kDa) and OMP J2 (16 kDa), were identified with similar gene sequences.
- Genes for OMP J1 and OMP J2 were found in 100% of tested M. catarrhalis isolates.
- OMP J1/J2 presence was significantly associated with specific genetic lineages and complement resistance (P < 0.01).
- DeltaompJ2 mutants showed reduced clearance from mouse lungs compared to wild-type, particularly the complement-resistant strain (P < 0.001).
Conclusions:
- Moraxella catarrhalis possesses novel outer membrane proteins (OMP J1 and OMP J2) with variant forms.
- These proteins are conserved across diverse isolates and linked to specific genetic traits.
- OMP J1 and OMP J2 contribute to bacterial survival and clearance from the lungs, suggesting a role in pathogenesis.
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