The Essential Protein for Bacterial Flagella Formation FlgJ Functions as a β-N-Acetylglucosaminidase

Herlihey, Francesca A.; Moynihan, Patrick J.; Clarke, Anthony J.

doi:10.1074/jbc.m114.603944

Cited by 60 publications

(74 citation statements)

References 54 publications

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“…Recently, a PG-lytic enzyme associated with the flagella, FlgJ, was shown to cleave peptidoglycan between GlcNAc and MurNAc saccharides, acting as an endospecific N-acetylglucosaminidase (56). Another study showed that Helicobacter pylori and Salmonella typhimurium required activity of housekeeping lytic transglycosylases for functionality of the flagella (i.e.…”

Section: Discussionmentioning

confidence: 99%

“…motility), but not flagella assembly (57). This work proposed a model in which the N-actelyglucosaminidase activity of FlgJ clears peptidoglycan to facilitate flagella assembly, and subsequently housekeeping (involved in cell growth and cell wall maintenance) lytic transglycosylases remodel the opening in the peptidoglycan, producing 1,6-anhydromuramoyl peptidoglycan ends that interact with MotB, the flagellar motor protein (56,57). Does the difference in structure (and likely activity) between type III-associated PG-lytic enzyme EtgA and the flagellar FlgJ somehow play a nuanced role in assembly of each system, or does it reflect a different evolutionary acquirement of a PG-lytic enzyme?…”

Section: Discussionmentioning

confidence: 99%

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Structural Analysis of a Specialized Type III Secretion System Peptidoglycan-cleaving Enzyme

Burkinshaw

Deng

Lameignère

et al. 2015

Journal of Biological Chemistry

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Background: Bacterial virulence-associated type III secretion system (T3SS) assembly requires a dedicated enzyme to penetrate peptidoglycan (PG). Results: We structurally characterized a T3SS PG-lytic enzyme, identified catalytically important residues, and characterized its activity. Conclusion:The active site is similar to lysozymes and lytic transglycosylases and interaction with the T3SS enhances activity. Significance: Structural information is critical for development of drugs targeting T3SS PG-lytic enzymes.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Structural Analysis of a Specialized Type III Secretion System Peptidoglycan-cleaving Enzyme

Burkinshaw

Deng

Lameignère

et al. 2015

Journal of Biological Chemistry

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“…Reaction specificity of SltF. The heavy water assay previously developed by Herlihey et al (6) was used to identify the type of lytic activity that SltF from R. sphaeroides catalyzes. S. Typhimurium PG in 50 mM sodium phosphate buffer (pH 7.0), prepared in […”

Section: Resultsmentioning

confidence: 99%

“…During flagellum assembly, extensive modifications need to occur to the peptidoglycan (PG) sacculus to accommodate the insertion of the secretion apparatus, as well as to stabilize the function of this system by acting as an assembly scaffold (6,7). PG is a heteropolymer of glycan strands and peptide chains forming a rigid network (sacculus) that completely surrounds bacterial cells to maintain the integrity of their cytoplasmic membranes.…”

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confidence: 99%

“…FlgJ of the flagellar operon has been shown to be required for the proper assembly of the flagellum. The FlgJs produced by the beta-and gammaproteobacteria, such as Salmonella enterica serovar Typhimurium, are bifunctional enzymes, possessing an N-terminal domain responsible for proper rod assembly (13)(14)(15)(16)(17)(18)) and a C-terminal domain possessing ␤-Nacetylglucosaminidase activity (6). Various alphaproteobacteria, including R. sphaeroides, possess FlgJ homologues that lack the C-terminal PG lytic domain.…”

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confidence: 99%

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Modulation of the Lytic Activity of the Dedicated Autolysin for Flagellum Formation SltF by Flagellar Rod Proteins FlgB and FlgF

et al. 2016

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SltF was identified previously as an autolysin required for the assembly of flagella in the alphaproteobacteria, but the nature of its peptidoglycan lytic activity remained unknown. Sequence alignment analyses suggest that it could function as either a muramidase, lytic transglycosylase, or ␤-N-acetylglucosaminidase. Recombinant SltF from Rhodobacter sphaeroides was purified to apparent homogeneity, and it was demonstrated to function as a lytic transglycosylase based on enzymatic assays involving mass spectrometric analyses. Circular dichroism (CD) analysis determined that it is composed of 83.4% ␣-structure and 1.48% ␤-structure and thus is similar to family 1A lytic transglycosylases. However, alignment of apparent SltF homologs identified in the genome database defined a new subfamily of the family 1 lytic transglycosylases. SltF was demonstrated to be endo-acting, cleaving within chains of peptidoglycan, with optimal activity at pH 7.0. Its activity is modulated by two flagellar rod proteins, FlgB and FlgF: FlgB both stabilizes and stimulates SltF activity, while FlgF inhibits it. Invariant Glu57 was confirmed as the sole catalytic acid/base residue of SltF. IMPORTANCEThe bacterial flagellum is comprised of a basal body, hook, and helical filament, which are connected by a rod structure. With a diameter of approximately 4 nm, the rod is larger than the estimated pore size within the peptidoglycan sacculus, and hence its insertion requires the localized and controlled lysis of this essential cell wall component. In many beta-and gammaproteobacteria, this lysis is catalyzed by the ␤-N-acetylglucosaminidase domain of FlgJ. However, FlgJ of the alphaproteobacteria lacks this activity and instead it recruits a separate enzyme, SltF, for this purpose. In this study, we demonstrate that SltF functions as a newly identified class of lytic transglycosylases and that its autolytic activity is uniquely modulated by two rod proteins, FlgB and FlgF.T he photosynthetic bacterium Rhodobacter sphaeroides is motile through the use of a single, subpolar flagellum (1). This locomotive organelle is tightly regulated and comprised of approximately 25 different proteins arranged into three major substructures: a basal body, hook, and thin helical filament. The basal body spans the bacterial cell envelope and contains the motor, a flagellum-specific type III export apparatus and at least four ringlike structures which are all connected by a filamentous rod (2). The rod assembles into a proximal rod that lies between the MS ring and the cell wall, which is composed of nine subunits of FliE (3) and six subunits of FlgB, FlgC, and FlgF (4), as well as a distal rod that is composed of 26 subunits of FlgG (5).During flagellum assembly, extensive modifications need to occur to the peptidoglycan (PG) sacculus to accommodate the insertion of the secretion apparatus, as well as to stabilize the function of this system by acting as an assembly scaffold (6, 7). PG is a heteropolymer of glycan strands and peptide chains forming a rig...

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Muropeptides in Pseudomonas aeruginosa and their Role as Elicitors of β‐Lactam‐Antibiotic Resistance

et al. 2016

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In Abbildung 2B dieser Zuschrift wurden die Produkte der Reaktionen von 2e mit PBP4 und AmpDh3 verwechselt. Die Reaktion von 2e mit PBP4 führt zu 2d,und die Reaktion von 2e mit AmpDh3 zu 2a,w ie in der korrigierten Abbildung 2unten gezeigt. In Haupttext und Abbildungslegende ergeben sich dadurch keine ¾nderungen. Figure 2. A) Chemical structures of detected muropeptides. B) The chemoenzymatic syntheses of six muropeptides.

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The Essential Protein for Bacterial Flagella Formation FlgJ Functions as a β-N-Acetylglucosaminidase

Cited by 60 publications

References 54 publications

Structural Analysis of a Specialized Type III Secretion System Peptidoglycan-cleaving Enzyme

Structural Analysis of a Specialized Type III Secretion System Peptidoglycan-cleaving Enzyme

Modulation of the Lytic Activity of the Dedicated Autolysin for Flagellum Formation SltF by Flagellar Rod Proteins FlgB and FlgF

Muropeptides in Pseudomonas aeruginosa and their Role as Elicitors of β‐Lactam‐Antibiotic Resistance

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