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Title: Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus

Abstract

ABSTRACT Bacterial pathogensNeisseria meningitidisandBrucella abortuspose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acuteN. meningitidisinfections remains high despite antibiotics, and brucellosis presents alimentary and health consequences. Superoxide dismutases are master regulators of reactive oxygen and general pathogenicity factors and are therefore therapeutic targets. Cu,Zn superoxide dismutases (SODs) localized to the periplasm promote survival by detoxifying superoxide radicals generated by major host antimicrobial immune responses. We discovered that passive immunization with an antibody directed atN. meningitidisSOD (NmSOD) was protective in a mouse infection model. To define the relevant atomic details and solution assembly states of this important virulence factor, we report high-resolution and X-ray scattering analyses of NmSOD and of SOD fromB. abortus(BaSOD). The NmSOD structures revealed an auxiliary tetrahedral Cu-binding site bridging the dimer interface; mutational analyses suggested that this metal site contributes to protein stability, with implications for bacterial defense mechanisms. Biochemical and structural analyses informed us about electrostatic substrate guidance, dimer assembly, and an exposed C-terminal epitope in the NmSOD dimer. In contrast, the monomeric BaSOD structure provided insights for extending immunogenic peptide epitopes derived from the protein. These collective results reveal unique contributions of SOD to pathogenic virulence, refine predictivemore » motifs for distinguishing SOD classes, and suggest general targets for antibacterial immune responses. The identified functional contributions, motifs, and targets distinguishing bacterial and eukaryotic SOD assemblies presented here provide a foundation for efforts to develop SOD-specific inhibitors of or vaccines against these harmful pathogens. IMPORTANCEBy protecting microbes against reactive oxygen insults, SODs aid survival of many bacteria within their hosts. Despite the ubiquity and conservation of these key enzymes, notable species-specific differences relevant to pathogenesis remain undefined. To probe mechanisms that govern the functioning ofNeisseria meningitidisandBrucella abortusSODs, we used X-ray structures, enzymology, modeling, and murine infection experiments. We identified virulence determinants common to the two homologs, assembly differences, and a unique metal reservoir within meningococcal SOD that stabilizes the enzyme and may provide a safeguard against copper toxicity. The insights reported here provide a rationale and a basis for SOD-specific drug design and an extension of immunogen design to target two important pathogens that continue to pose global health threats.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1407285
DOE Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article
Journal Name:
Journal of Bacteriology
Additional Journal Information:
Journal Volume: 197; Journal Issue: 24; Journal ID: ISSN 0021-9193
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEAR CHEMISTRY

Citation Formats

Pratt, Ashley J., DiDonato, Michael, Shin, David S., Cabelli, Diane E., Bruns, Cami K., Belzer, Carol A., Gorringe, Andrew R., Langford, Paul R., Tabatabai, Louisa B., Kroll, J. Simon, Tainer, John A., Getzoff, Elizabeth D., and Stock, A. M.. Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus. United States: N. p., 2015. Web. doi:10.1128/JB.00343-15.
Pratt, Ashley J., DiDonato, Michael, Shin, David S., Cabelli, Diane E., Bruns, Cami K., Belzer, Carol A., Gorringe, Andrew R., Langford, Paul R., Tabatabai, Louisa B., Kroll, J. Simon, Tainer, John A., Getzoff, Elizabeth D., & Stock, A. M.. Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus. United States. https://doi.org/10.1128/JB.00343-15
Pratt, Ashley J., DiDonato, Michael, Shin, David S., Cabelli, Diane E., Bruns, Cami K., Belzer, Carol A., Gorringe, Andrew R., Langford, Paul R., Tabatabai, Louisa B., Kroll, J. Simon, Tainer, John A., Getzoff, Elizabeth D., and Stock, A. M.. Mon . "Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus". United States. https://doi.org/10.1128/JB.00343-15. https://www.osti.gov/servlets/purl/1407285.
@article{osti_1407285,
title = {Structural, Functional, and Immunogenic Insights on Cu,Zn Superoxide Dismutase Pathogenic Virulence Factors from Neisseria meningitidis and Brucella abortus},
author = {Pratt, Ashley J. and DiDonato, Michael and Shin, David S. and Cabelli, Diane E. and Bruns, Cami K. and Belzer, Carol A. and Gorringe, Andrew R. and Langford, Paul R. and Tabatabai, Louisa B. and Kroll, J. Simon and Tainer, John A. and Getzoff, Elizabeth D. and Stock, A. M.},
abstractNote = {ABSTRACT Bacterial pathogensNeisseria meningitidisandBrucella abortuspose threats to human and animal health worldwide, causing meningococcal disease and brucellosis, respectively. Mortality from acuteN. meningitidisinfections remains high despite antibiotics, and brucellosis presents alimentary and health consequences. Superoxide dismutases are master regulators of reactive oxygen and general pathogenicity factors and are therefore therapeutic targets. Cu,Zn superoxide dismutases (SODs) localized to the periplasm promote survival by detoxifying superoxide radicals generated by major host antimicrobial immune responses. We discovered that passive immunization with an antibody directed atN. meningitidisSOD (NmSOD) was protective in a mouse infection model. To define the relevant atomic details and solution assembly states of this important virulence factor, we report high-resolution and X-ray scattering analyses of NmSOD and of SOD fromB. abortus(BaSOD). The NmSOD structures revealed an auxiliary tetrahedral Cu-binding site bridging the dimer interface; mutational analyses suggested that this metal site contributes to protein stability, with implications for bacterial defense mechanisms. Biochemical and structural analyses informed us about electrostatic substrate guidance, dimer assembly, and an exposed C-terminal epitope in the NmSOD dimer. In contrast, the monomeric BaSOD structure provided insights for extending immunogenic peptide epitopes derived from the protein. These collective results reveal unique contributions of SOD to pathogenic virulence, refine predictive motifs for distinguishing SOD classes, and suggest general targets for antibacterial immune responses. The identified functional contributions, motifs, and targets distinguishing bacterial and eukaryotic SOD assemblies presented here provide a foundation for efforts to develop SOD-specific inhibitors of or vaccines against these harmful pathogens. IMPORTANCEBy protecting microbes against reactive oxygen insults, SODs aid survival of many bacteria within their hosts. Despite the ubiquity and conservation of these key enzymes, notable species-specific differences relevant to pathogenesis remain undefined. To probe mechanisms that govern the functioning ofNeisseria meningitidisandBrucella abortusSODs, we used X-ray structures, enzymology, modeling, and murine infection experiments. We identified virulence determinants common to the two homologs, assembly differences, and a unique metal reservoir within meningococcal SOD that stabilizes the enzyme and may provide a safeguard against copper toxicity. The insights reported here provide a rationale and a basis for SOD-specific drug design and an extension of immunogen design to target two important pathogens that continue to pose global health threats.},
doi = {10.1128/JB.00343-15},
url = {https://www.osti.gov/biblio/1407285}, journal = {Journal of Bacteriology},
issn = {0021-9193},
number = 24,
volume = 197,
place = {United States},
year = {2015},
month = {10}
}

Works referenced in this record:

Novel dimeric interface and electrostatic recognition in bacterial Cu,Zn superoxide dismutase
journal, November 1996


DAMMIF, a program for rapid ab-initio shape determination in small-angle scattering
journal, January 2009


Cloning, expression, and occurrence of the Brucella Cu-Zn superoxide dismutase.
journal, January 1990


The Structure of Human Extracellular Copper–Zinc Superoxide Dismutase at 1.7 Å Resolution: Insights into Heparin and Collagen Binding
journal, May 2009


Amyloid-like filaments and water-filled nanotubes formed by SOD1 mutant proteins linked to familial ALS
journal, May 2003


Single mutations at the subunit interface modulate copper reactivity in Photobacterium leiognathi Cu,Zn superoxide dismutase
journal, May 2001


The role of arginine 143 in the electrostatics and mechanism of Cu, Zn superoxide dismutase: Computational and experimental evaluation by mutational analysis
journal, May 1994


The reactivity of anti-peptide antibodies is a function of the atomic mobility of sites in a protein
journal, November 1984


Active-site Copper and Zinc Ions Modulate the Quaternary Structure of Prokaryotic Cu,Zn Superoxide Dismutase
journal, March 2003


Super-Resolution in Solution X-Ray Scattering and Its Applications to Structural Systems Biology
journal, May 2013


Copper at the Front Line of the Host-Pathogen Battle
journal, September 2012


Candida albicans SOD5 represents the prototype of an unprecedented class of Cu-only superoxide dismutases required for pathogen defense
journal, April 2014


The role of innate immune signals in immunity to Brucella abortus
journal, January 2012


The Good and the Bad of Antibiotics
journal, July 2013


History of biological metal utilization inferred through phylogenomic analysis of protein structures
journal, May 2010


UCSF Chimera?A visualization system for exploratory research and analysis
journal, January 2004


Treatment strategies for central nervous system infections: an update
journal, October 2014


Superoxide Stress Decreases Expression of srfA through Inhibition of Transcription of the comQXP Quorum-Sensing Locus in Bacillus subtilis
journal, February 2006


A vaccine against serogroup B Neisseria meningitidis: dealing with uncertainty
journal, May 2014


AMoRe : an automated package for molecular replacement
journal, March 1994


Brucella as a Potential Agent of Bioterrorism
journal, February 2013


PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010


The changing and dynamic epidemiology of meningococcal disease
journal, May 2012


Detection and Quantification of Superoxide Formed within the Periplasm of Escherichia coli
journal, August 2006


Current Perspectives on Rabies Postexposure Prophylaxis
journal, April 2015


Faster superoxide dismutase mutants designed by enhancing electrostatic guidance
journal, July 1992


Mechanisms of Oxidative Protein Folding in the Bacterial Cell Envelope
journal, October 2010


A protein isolated from Brucella abortus is a copper-zinc superoxide dismutase
journal, January 1990


Physiological Roles of Mitochondrial Reactive Oxygen Species
journal, October 2012


Structural Characterization of the Active Site of Brucella abortus Cu-Zn Superoxide Dismutase: A 15N and 1H NMR Investigation
journal, September 1995


Accurate assessment of mass, models and resolution by small-angle scattering
journal, April 2013


The structural biochemistry of the superoxide dismutases
journal, February 2010


SOD2 to SOD1 Switch in Breast Cancer
journal, January 2014


Determination and analysis of the 2 Å structure of copper, zinc superoxide dismutase
journal, September 1982


Bacterial species exhibit diversity in their mechanisms and capacity for protein disulfide bond formation
journal, August 2008


Uniqueness of ab initio shape determination in small-angle scattering
journal, April 2003


Clinical Manifestations of Human Brucellosis: A Systematic Review and Meta-Analysis
journal, December 2012


Bacterial redox sensors
journal, December 2004


Assembly and antigenicity of the Neisseria gonorrhoeae pilus mapped with antibodies.
journal, January 1996


On the possible roles of N-terminal His-rich domains of Cu,Zn SODs of some Gram-negative bacteria
journal, January 2012


The new global map of human brucellosis
journal, February 2006


Structure and mechanism of copper, zinc superoxide dismutase
journal, November 1983


Evolution of CuZn superoxide dismutase and the Greek Key β-barrel structural motif
journal, January 1989


PRIMUS: a Windows PC-based system for small-angle scattering data analysis
journal, September 2003


Crystallography & NMR System: A New Software Suite for Macromolecular Structure Determination
journal, September 1998


Conquering the Meningococcus
journal, January 2007


Function of the Greek key connection analysed using circular permutants of superoxide dismutase
journal, May 1997


ElNemo: a normal mode web server for protein movement analysis and the generation of templates for molecular replacement
journal, July 2004


Chlorovirus PBCV-1 Encodes an Active Copper-Zinc Superoxide Dismutase
journal, August 2014


A Histidine-rich Metal Binding Domain at the N Terminus of Cu,Zn-Superoxide Dismutases from Pathogenic Bacteria: A NOVEL STRATEGY FOR METAL CHAPERONING
journal, May 2001


A History of the Development of Brucella Vaccines
journal, January 2013


Aggregation propensities of superoxide dismutase G93 hotspot mutants mirror ALS clinical phenotypes
journal, October 2014


A prokaryotic superoxide dismutase paralog lacking two Cu ligands: From largely unstructured in solution to ordered in the crystal
journal, May 2005


Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS)
journal, July 2009


Selenocysteine's mechanism of incorporation and evolution revealed in cDNAs of three glutathione peroxidases
journal, January 1988


Periplasmic Superoxide Dismutase in Meningococcal Pathogenicity
journal, January 1998


Meningococcal carriage and disease—Population biology and evolution
journal, June 2009


Progress in Brucella vaccine development
journal, March 2012


GNOM – a program package for small-angle scattering data processing
journal, October 1991


Towards a Brucella vaccine for humans
journal, May 2010


Antimicrobial Resistance and Virulence: a Successful or Deleterious Association in the Bacterial World?
journal, April 2013


A Novel Heme Protein, the Cu,Zn-Superoxide Dismutase from Haemophilus ducreyi
journal, May 2001


Cu, Zn superoxide dismutase structure from a microbial pathogen establishes a class with a conserved dimer interface 1 1Edited by D. C. Rees
journal, February 2000


Bacterial manipulation of innate immunity to promote infection
journal, February 2010


ALS Mutants of Human Superoxide Dismutase Form Fibrous Aggregates Via Framework Destabilization
journal, September 2003


Advances in treatment of bacterial meningitis
journal, November 2012


FoXS: a web server for rapid computation and fitting of SAXS profiles
journal, May 2010


Functional and crystallographic characterization of Salmonella typhimurium Cu,Zn superoxide dismutase coded by the sodCI virulence gene 1 1Edited by R. Huber
journal, September 2000


Electrostatic recognition between superoxide and copper, zinc superoxide dismutase
journal, November 1983


Comprehensive macromolecular conformations mapped by quantitative SAXS analyses
journal, April 2013


Virulent Salmonella typhimurium has two periplasmic Cu, Zn-superoxide dismutases
journal, June 1999


Adverse events in humans associated with accidental exposure to the livestock brucellosis vaccine RB51
journal, September 2004


In vivo copper-mediated free radical production: an ESR spin-trapping study
journal, April 2002


Structural Basis of Heme Binding in the Cu,Zn Superoxide Dismutase from Haemophilus ducreyi
journal, February 2009


XtalView/Xfit—A Versatile Program for Manipulating Atomic Coordinates and Electron Density
journal, April 1999


Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH-oxidase and nitric oxide synthase
journal, December 1997


Unique structural features of the monomeric Cu,Zn superoxide dismutase from Escherichia coli, revealed by X-ray crystallography
journal, December 1997


Direct Oxidation of the [2Fe-2S] Cluster in SoxR Protein by Superoxide: DISTINCT DIFFERENTIAL SENSITIVITY TO SUPEROXIDE-MEDIATED SIGNAL TRANSDUCTION
journal, August 2012


Conserved Enzyme-Substrate Electrostatic Attraction in Prokaryotic Cu,Zn Superoxide Dismutases
journal, March 1998


Atomic structures of wild-type and thermostable mutant recombinant human Cu,Zn superoxide dismutase.
journal, July 1992


Evolutionary constraints for dimer formation in prokaryotic Cu,Zn superoxide dismutase 1 1Edited by R. Huber
journal, January 1999


Reconciling the chemistry and biology of reactive oxygen species
journal, April 2008


Microbial metalloproteomes are largely uncharacterized
journal, July 2010


Neisseria meningitidis factor H-binding protein fHbp: a key virulence factor and vaccine antigen
journal, February 2015


Human copper-zinc superoxide dismutase complements superoxide dismutase-deficient Escherichia coli mutants.
journal, October 1987


Genetically Engineered Polymers of Human CuZn Superoxide Dismutase
journal, March 1989


    Works referencing / citing this record:

    Immune Response to Mucosal Brucella Infection
    journal, August 2019


    Antigenicity of Brucella Proteins by the Elisa test
    journal, March 2019