Document Detail


Structural and functional characterization of the Streptococcus pneumoniae RrgB pilus backbone D1 domain.
MedLine Citation:
PMID:  21367860     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Maria Antonietta Gentile; Sara Melchiorre; Carla Emolo; Monica Moschioni; Claudia Gianfaldoni; Laura Pancotto; Ilaria Ferlenghi; Maria Scarselli; Werner Pansegrau; Daniele Veggi; Marcello Merola; Francesca Cantini; Paolo Ruggiero; Lucia Banci; Vega Masignani
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-03-02
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  286     ISSN:  1083-351X     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2011 Apr 
Date Detail:
Created Date:  2011-04-18     Completed Date:  2011-06-28     Revised Date:  2012-09-19    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  14588-97     Citation Subset:  IM    
Affiliation:
Novartis Vaccines and Diagnostics Research Center, Via Fiorentina 1, Siena 53100, Italy.
Data Bank Information
Bank Name/Acc. No.:
PDB/2L40
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MeSH Terms
Descriptor/Qualifier:
Animals
Bacterial Proteins / chemistry
Cell Adhesion
Disease Models, Animal
Epitopes / chemistry
Fimbriae Proteins / chemistry*,  genetics
Genetic Complementation Test
Magnetic Resonance Spectroscopy / methods
Mice
Mice, Inbred BALB C
Mutagenesis, Site-Directed
Peptides / chemistry
Protein Conformation
Protein Structure, Tertiary
Sepsis / metabolism
Streptococcus pneumoniae / metabolism*
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Epitopes; 0/Peptides; 147680-16-8/Fimbriae Proteins
Comments/Corrections

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