Crystal Structure of Neisserial Surface Protein A (NspA), a Conserved Outer Membrane Protein with Vaccine Potential*

The neisserial surface protein A (NspA) from Neisseria meningitidis is a promising vaccine candidate because it is highly conserved among meningococcal strains and induces bactericidal antibodies. NspA is a homolog of the Opa proteins, which mediate adhesion to host cells. Here, we present the crystal structure of NspA, determined to 2.55-Å resolution. NspA forms an eight-stranded antiparallel β-barrel. The four loops at the extracellular side of the NspA molecule form a long cleft, which contains mainly hydrophobic residues and harbors a detergent molecule, suggesting that the protein might function in the binding of hydrophobic ligands, such as lipids. In addition, the structure provides a starting point for structure-based vaccine design.

[1]  M. Achtman,et al.  Sequence Diversity, Predicted Two-Dimensional Protein Structure, and Epitope Mapping of Neisserial Opa Proteins , 1998, Journal of bacteriology.

[2]  Z. Otwinowski,et al.  [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[3]  M. Achtman,et al.  Crystal structure of the OpcA integral membrane adhesin from Neisseria meningitidis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  D. Ferguson,et al.  Carcinoembryonic antigens (CD66) on epithelial cells and neutrophils are receptors for Opa proteins of pathogenic neisseriae , 1996, Molecular microbiology.

[5]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[6]  K. Tanizawa,et al.  Structure of a quinohemoprotein amine dehydrogenase with an uncommon redox cofactor and highly unusual crosslinking , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. Mandrell,et al.  Importance of complement source in bactericidal activity of human antibody and murine monoclonal antibody to meningococcal group B polysaccharide , 1983, Infection and immunity.

[8]  G. Schulz,et al.  Structure of the outer membrane protein A transmembrane domain , 1998, Nature Structural Biology.

[9]  K. Diederichs,et al.  Crystal structure of Omp32, the anion-selective porin from Comamonas acidovorans, in complex with a periplasmic peptide at 2.1 A resolution. , 2000, Structure.

[10]  G. Schulz.  beta-Barrel membrane proteins. , 2000, Current opinion in structural biology.

[11]  G. Schulz.  The structure of bacterial outer membrane proteins. , 2002, Biochimica et biophysica acta.

[12]  G. Schulz.  β-Barrel membrane proteins , 2000 .

[13]  E A Merritt,et al.  Raster3D Version 2.0. A program for photorealistic molecular graphics. , 1994, Acta crystallographica. Section D, Biological crystallography.

[14]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[15]  Wing-Yiu Choy,et al.  Solution structure and dynamics of the outer membrane enzyme PagP by NMR , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Tommassen,et al.  In vitro folding of Escherichia coli outer-membrane phospholipase A. , 1995, European journal of biochemistry.

[17]  M. Leinonen,et al.  Protective efficacy of monoclonal antibodies to class 1 and class 3 outer membrane proteins of Neisseria meningitidis B:15:P1.16 in infant rat infection model: new prospects for vaccine development. , 1987, Microbial pathogenesis.

[18]  A. Lucas,et al.  Functional Activity of Anti-Neisserial Surface Protein A Monoclonal Antibodies against Strains of Neisseria meningitidis Serogroup B , 2001, Infection and Immunity.

[19]  B. Dijkstra,et al.  Bacterial phospholipase A: structure and function of an integral membrane phospholipase. , 2000, Biochimica et biophysica acta.

[20]  R M Esnouf,et al.  Further additions to MolScript version 1.4, including reading and contouring of electron-density maps. , 1999, Acta crystallographica. Section D, Biological crystallography.

[21]  R. Mandrell,et al.  Measurement of antibodies to meningococcal group B polysaccharide: low avidity binding and equilibrium binding constants. , 1982, Journal of immunology.

[22]  J. Tommassen,et al.  The ultimate localization of an outer membrane protein of Escherichia coli K‐12 is not determined by the signal sequence. , 1983, The EMBO journal.

[23]  D. Granoff,et al.  Sequential Immunization with Vesicles Prepared from Heterologous Neisseria meningitidis Strains Elicits Broadly Protective Serum Antibodies to Group B Strains , 2002, Infection and Immunity.

[24]  C. Frasch.  Vaccines for prevention of meningococcal disease , 1989, Clinical Microbiology Reviews.

[25]  N. Cadieux,et al.  Bactericidal and Cross-Protective Activities of a Monoclonal Antibody Directed against Neisseria meningitidis NspA Outer Membrane Protein , 1999, Infection and Immunity.

[26]  R Miller,et al.  Optimizing Shake-and-Bake for proteins. , 1999, Acta crystallographica. Section D, Biological crystallography.

[27]  N. Cadieux,et al.  Antigenic and Molecular Conservation of the Gonococcal NspA Protein , 1999, Infection and Immunity.

[28]  C. Sander,et al.  Protein structure comparison by alignment of distance matrices. , 1993, Journal of molecular biology.

[29]  P Gros,et al.  Crystal structure of the outer membrane protease OmpT from Escherichia coli suggests a novel catalytic site , 2001, The EMBO journal.

[30]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[31]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[32]  R. Belshe,et al.  Safety and immunogenicity of meningococcal A and C polysaccharide conjugate vaccine in adults , 1994, Infection and immunity.

[33]  E. Gotschlich,et al.  CGM1a antigen of neutrophils, a receptor of gonococcal opacity proteins. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[34]  K. Wilson,et al.  Efficient anisotropic refinement of macromolecular structures using FFT. , 1999, Acta crystallographica. Section D, Biological crystallography.

[35]  Alexandre M J J Bonvin,et al.  Immunogenicity of peptide-vaccine candidates predicted by molecular dynamics simulations. , 2003, Journal of molecular biology.

[36]  G. Schulz,et al.  The structure of the outer membrane protein OmpX from Escherichia coli reveals possible mechanisms of virulence. , 1999, Structure.

[37]  M. Achtman,et al.  The 5C protein of Neisseria meningitidis is highly immunogenic in humans and induces bactericidal antibodies. , 1993, The Journal of infectious diseases.

[38]  J. Poolman,et al.  Alternative vaccine strategies to prevent serogroup B meningococcal diseases. , 2001, Vaccine.

[39]  D. Caugant,et al.  A new variant of serosubtype P1.16 in Neisseria meningitidis from Norway, associated with increased resistance to bactericidal antibodies induced by a serogroup B outer membrane protein vaccine. , 1993, Microbial pathogenesis.

[40]  R. Rappuoli,et al.  Development and phase 1 clinical testing of a conjugate vaccine against meningococcus A and C. , 1992, Vaccine.

[41]  D. Granoff,et al.  Differences in Surface Expression of NspA amongNeisseria meningitidis Group B Strains , 1999, Infection and Immunity.