Streptococcus pneumoniae expresses on its surface adhesive pili, involved in bacterial attachment to epithelial cells and virulence. The pneumococcal pilus is composed of three proteins, RrgA, RrgB, and RrgC, each stabilized by intramolecular isopeptide bonds and covalently polymerized by means of intermolecular isopeptide bonds to form an extended fiber. RrgB is the pilus scaffold subunit and is protective in vivo in mouse models of sepsis and pneumonia, thus representing a potential vaccine candidate. The crystal structure of a major RrgB C-terminal portion featured an organization into three independently folded protein domains (D2-D4), whereas the N-terminal D1 domain (D1) remained unsolved. We have tested the four single recombinant RrgB domains in active and passive immunization studies and show that D1 is the most effective, providing a level of protection comparable with that of the full-length protein. To elucidate the structural features of D1, we solved the solution structure of the recombinant domain by NMR spectroscopy. The spectra analysis revealed that D1 has many flexible regions, does not contain any intramolecular isopeptide bond, and shares with the other domains an Ig-like fold. In addition, we demonstrated, by site-directed mutagenesis and complementation in S. pneumoniae, that the D1 domain contains the Lys residue (Lys-183) involved in the formation of the intermolecular isopeptide bonds and pilus polymerization. Finally, we present a model of the RrgB protein architecture along with the mapping of two surface-exposed linear epitopes recognized by protective antisera.
机构:
CEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
CNRS, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Univ Grenoble 1, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, FranceCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
El Mortaji, Lamya
Contreras-Martel, Carlos
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机构:
CEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
CNRS, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Univ Grenoble 1, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, FranceCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Contreras-Martel, Carlos
Moschioni, Monica
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机构:
Novartis Vaccines & Diagnost, I-53100 Siena, ItalyCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Moschioni, Monica
Ferlenghi, Ilaria
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Novartis Vaccines & Diagnost, I-53100 Siena, ItalyCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Ferlenghi, Ilaria
Manzano, Clothilde
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机构:
CEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
CNRS, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Univ Grenoble 1, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, FranceCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Manzano, Clothilde
Vernet, Thierry
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机构:
CEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
CNRS, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Univ Grenoble 1, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, FranceCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Vernet, Thierry
Dessen, Andrea
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CEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
CNRS, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Univ Grenoble 1, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, FranceCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Dessen, Andrea
Di Guilmi, Anne Marie
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机构:
CEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
CNRS, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
Univ Grenoble 1, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, FranceCEA, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble, France
机构:
Emory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Dept Vet Affairs Med Ctr Atlanta, Decatur, GA 30033 USAEmory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Zaehner, Dorothea
Gandhi, Ashish R.
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Emory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USAEmory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Gandhi, Ashish R.
Stuchlik, Olga
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机构:
Ctr Dis Control & Prevent, Biotechnol Core Facil Branch, Atlanta, GA 30329 USAEmory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Stuchlik, Olga
Reed, Matthew
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Ctr Dis Control & Prevent, Biotechnol Core Facil Branch, Atlanta, GA 30329 USAEmory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Reed, Matthew
Pohl, Jan
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Ctr Dis Control & Prevent, Biotechnol Core Facil Branch, Atlanta, GA 30329 USAEmory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Pohl, Jan
Stephens, David S.
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Emory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
Emory Univ, Sch Med, Rollins Res Ctr, Dept Microbiol & Immunol, Atlanta, GA 30322 USA
Dept Vet Affairs Med Ctr Atlanta, Decatur, GA 30033 USAEmory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA 30322 USA
机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
Izore, Thierry
Contreras-Martel, Carlos
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机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
Contreras-Martel, Carlos
El Mortaji, Lamya
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机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
El Mortaji, Lamya
Manzano, Clothilde
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h-index: 0
机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
Manzano, Clothilde
Terrasse, Remy
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机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
Terrasse, Remy
Vernet, Thierry
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机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
Vernet, Thierry
Di Guilmi, Anne Marie
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h-index: 0
机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France
Di Guilmi, Anne Marie
Dessen, Andrea
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h-index: 0
机构:
CEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, FranceCEA CNRS UJF, Inst Biol Struct Jean Pierre Ebel, UMR 5075, F-38027 Grenoble, France