Favini-Stabile S.,CNRS Institute of Pharmacology and Structural Biology |
Favini-Stabile S.,CEA Grenoble |
Favini-Stabile S.,French National Center for Scientific Research |
Contreras-Martel C.,CNRS Institute of Pharmacology and Structural Biology |
And 9 more authors.
Environmental Microbiology | Year: 2013
Peptidoglycan is a major determinant of cell shape in bacteria, and its biosynthesis involves the concerted action of cytoplasmic, membrane-associated and periplasmic enzymes. Within the cytoplasm, Mur enzymes catalyse the first steps leading to peptidoglycan precursor biosynthesis, and have been suggested as being part of a multicomponent complex that could also involve the transglycosylase MurG and the cytoskeletal protein MreB. In order to initialize the characterization of a potential Mur interaction network, we purified MurD, MurE, MurF, MurG and MreB from Thermotoga maritima and characterized their interactions using membrane blotting and surface plasmon resonance. MurD, MurE and MurF all recognize MurG and MreB, but not each other, while the two latter proteins interact. In addition, we solved the crystal structures of MurD, MurE and MurF, which indicate that their C-termini display high conformational flexibilities. The differences in Mur conformations could be important parameters for the stability of an intracytoplasmic murein biosynthesis complex. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd. Source
Tosi T.,CNRS Institute of Pharmacology and Structural Biology |
Tosi T.,French National Center for Scientific Research |
Tosi T.,CEA Grenoble |
Estrozi L.F.,CNRS Institute of Pharmacology and Structural Biology |
And 16 more authors.
Structure | Year: 2014
Secretins, the outer membrane components of several secretion systems in Gram-negative bacteria, assemble into channels that allow exoproteins to traverse the membrane. The membrane-inserted, multimeric regions of PscC, the Pseudomonas aeruginosa type III secretion system secretin, and PulD, the Klebsiella oxytoca type II secretion system secretin, were purified after cell-free synthesis and their structures analyzed by single particle cryoelectron microscopy. Both homomultimeric, barrel-like structures display a "cup and saucer" architecture. The "saucer" region of both secretins is composed of two distinct rings, with that of PulD being less segmented than that of PscC. Both secretins have a central chamber that is occluded by a plug linked to the chamber walls through hairpin-like structures. Comparisons with published structures from other bacterial systems reveal that secretins have regions of local structural flexibility, probably reflecting their evolved functions in protein secretion and needle assembly. © 2014 Elsevier Ltd. Source
Nikolaidis I.,CNRS Institute of Pharmacology and Structural Biology |
Nikolaidis I.,CEA Grenoble |
Nikolaidis I.,French National Center for Scientific Research |
Nikolaidis I.,University Utrecht |
And 7 more authors.
Protein Science | Year: 2014
Peptidoglycan is the main component of the bacterial cell wall. It is a complex, threedimensional mesh that surrounds the entire cell and is composed of strands of alternating glycan units crosslinked by short peptides. Its biosynthetic machinery has been, for the past five decades, a preferred target for the discovery of antibacterials. Synthesis of the peptidoglycan occurs sequentially within three cellular compartments (cytoplasm, membrane, and periplasm), and inhibitors of proteins that catalyze each stage have been identified, although not all are applicable for clinical use. A number of these antimicrobials, however, have been rendered inactive by resistance mechanisms. The employment of structural biology techniques has been instrumental in the understanding of such processes, as well as the development of strategies to overcome them. This review provides an overview of resistance mechanisms developed toward antibiotics that target bacterial cell wall precursors and its biosynthetic machinery. Strategies toward the development of novel inhibitors that could overcome resistance are also discussed. © 2014 The Protein Society. Source
Shaik M.M.,CNRS Institute of Pharmacology and Structural Biology |
Shaik M.M.,CEA Grenoble |
Shaik M.M.,French National Center for Scientific Research |
Maccagni A.,CNRS Institute of Pharmacology and Structural Biology |
And 12 more authors.
Journal of Biological Chemistry | Year: 2014
Pili are surface-attached, fibrous virulence factors that play key roles in the pathogenesis process of a number of bacterial agents. Streptococcus pneumoniae is a causative agent of pneumonia and meningitis, and the appearance of drug-resistance organisms has made its treatment challenging, especially in developing countries. Pneumococcus-expressed pili are composed of three structural proteins: RrgB, which forms the polymerized backbone, RrgA, the tip-associated adhesin, and RrgC, which presumably associates the pilus with the bacterial cell wall. Despite the fact that the structures of both RrgA and RrgB were known previously, structural information for RrgC was still lacking, impeding the analysis of a complete model of pilus architecture. Here, we report the structure of RrgC to 1.85 Å and reveal that it is a three-domain molecule stabilized by two intradomain isopeptide bonds. RrgC does not depend on pilus-specific sortases to become attached to the cell wall; instead, it binds the preformed pilus to the peptidoglycan by employing the catalytic activity of SrtA. A comprehensive model of the type 1 pilus from S. pneumoniae is also presented. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Source
Wong S.G.,CNRS Institute of Pharmacology and Structural Biology |
Wong S.G.,French National Center for Scientific Research |
Wong S.G.,CEA Grenoble |
Dessen A.,CNRS Institute of Pharmacology and Structural Biology |
And 3 more authors.
Nature Communications | Year: 2014
Alpha-2-macroglobulins (A2Ms) are plasma proteins that trap and inhibit a broad range of proteases and are major components of the eukaryotic innate immune system. Surprisingly, A2M-like proteins were identified in pathogenically invasive bacteria and species that colonize higher eukaryotes. Bacterial A2Ms are located in the periplasm where they are believed to provide protection to the cell by trapping external proteases through a covalent interaction with an activated thioester. Here we report the crystal structures and characterization of Salmonella enterica ser. Typhimurium A2M in different states of thioester activation. The structures reveal thirteen domains whose arrangement displays high similarity to proteins involved in eukaryotic immune defence. A structural lock mechanism maintains the stability of the buried thioester, a requirement for its protease-trapping activity. These findings indicate that bacteria have developed a rudimentary innate immune system whose mechanism mimics that of eukaryotes. © 2014 Macmillan Publishers Limited. All rights reserved. Source