1H, 13C and 15N resonance assignments of the C-terminal domain of HasB, a specific TonB like protein, from Serratia marcescens

Lefèvre, Julien; Simenel, Catherine; Delepelaire, Philippe; Delepierre, Murielle; Izadi-Pruneyre, Nadia

doi:10.1007/s12104-007-9055-7

Cited by 6 publications

(11 citation statements)

References 10 publications

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“…The solution structure of a longer construct (137 amino acids) was found to be monomeric [14], consistent with analytical-ultracentrifugation studies with similarly sized TonB constructs [12]. Recently the NMR assignments for the backbone of HasB CTD, a TonB-like protein from Serratia marcescens , were reported [15]. …”

Section: Introductionsupporting

confidence: 53%

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Molecular characterization of the TonB2 protein from the fish pathogenVibrio anguillarum

López

Peacock

Crosa

et al. 2009

Biochemical Journal

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In the fish pathogen Vibrio anguillarum the TonB2 protein is essential for the uptake of the indigenous siderophore anguibactin. Here we describe deletion mutants and alanine replacements affecting the final six amino acids of TonB2. Deletions of more than two amino acids of the TonB2 C-terminus abolished ferric-anguibactin transport, whereas replacement of the last three residues resulted in a protein with wild-type transport properties. We have solved the highresolution solution structure of the TonB2 C-terminal domain by NMR spectroscopy. The core of this domain (residues 121-206) has an αββαβ structure, whereas residues 76-120 are flexible and extended. This overall folding topology is similar to the Escherichia coli TonB C-terminal domain, albeit with two differences: the β4 strand found at the C-terminus of TonB is absent in TonB2, and loop 3 is extended by 9 Å (0.9 nm) in TonB2. By examining several mutants, we determined that a complete loop 3 is not essential for TonB2 activity. Our results indicate that the β4 strand of E. coli TonB is not required for activity of the TonB system across Gram-negative bacterial species. We have also determined, through NMR chemical-shift-perturbation experiments, that the E. coli TonB binds in vitro to the TonB box from the TonB2-dependent outer membrane transporter FatA; moreover, it can substitute in vivo for TonB2 during ferricanguibactin transport in V. anguillarum. Unexpectedly, TonB2 did not bind in vitro to the FatA TonB-box region, suggesting that additional factors may be required to promote this interaction. Overall our results indicate that TonB2 is a representative of a different class of TonB proteins.

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Section: Introductionsupporting

confidence: 53%

“…Details of the FhuA TonB box peptide have been described previously [15]. The FatA TonB box peptide has the sequence Glu-Ser-Ile-Thr-Val-Tyr-Gly-Glu-Ala, and the FhuA TonB-box peptide has the sequence Glu-Asp-Thr-Ile-Thr-Val-Thr-Ala-Ala-Pro.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of the TonB2 protein from the fish pathogenVibrio anguillarum

López

Peacock

Crosa

et al. 2009

Biochemical Journal

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“…The HasB 133 gene was amplified by PCR from plasmid pHasBpuc. 46 The reaction mixture included proofreading DNA polymerase Pfu (Stratagene) and each primer that we designed to incorporate 5′ EcoRI and 3′ HindIII sites for cloning the amplified product into the plasmid pBAD24. The sequence of the PCR forward primer was as follows: 5′-CAGGAGGAATTCACCATGAAGGTTCAGGAG-CAAAGCGT-3′.…”

Section: Strains and Plasmidsmentioning

confidence: 99%

“…46 Wildtype and mutant HasR, 29 HasA 47 and TonB 116 14 were expressed as previously described. HemRhis was expressed in LB medium at 30°C.…”

Section: Protein Expressionmentioning

confidence: 99%

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Modulation by Substrates of the Interaction between the HasR Outer Membrane Receptor and Its Specific TonB-like Protein, HasB

Lefèvre

Delepelaire

Delepierre

et al. 2008

Journal of Molecular Biology

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Structural and molecular determinants for the interaction of ExbB from Serratia marcescens and HasB, a TonB paralog

et al. 2022

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ExbB and ExbD are cytoplasmic membrane proteins that associate with TonB to convey the energy of the proton-motive force to outer membrane receptors in Gram-negative bacteria for iron uptake. The opportunistic pathogen Serratia marcescens (Sm) possesses both TonB and a heme-specific TonB paralog, HasB. ExbBSm has a long periplasmic extension absent in other bacteria such as E. coli (Ec). Long ExbB’s are found in several genera of Alphaproteobacteria, most often in correlation with a hasB gene. We investigated specificity determinants of ExbBSm and HasB. We determined the cryo-EM structures of ExbBSm and of the ExbB-ExbDSm complex from S. marcescens. ExbBSm alone is a stable pentamer, and its complex includes two ExbD monomers. We showed that ExbBSm extension interacts with HasB and is involved in heme acquisition and we identified key residues in the membrane domain of ExbBSm and ExbBEc, essential for function and likely involved in the interaction with TonB/HasB. Our results shed light on the class of inner membrane energy machinery formed by ExbB, ExbD and HasB.

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1H, 13C and 15N resonance assignments of the C-terminal domain of HasB, a specific TonB like protein, from Serratia marcescens

Cited by 6 publications

References 10 publications

Molecular characterization of the TonB2 protein from the fish pathogenVibrio anguillarum

Molecular characterization of the TonB2 protein from the fish pathogenVibrio anguillarum

Modulation by Substrates of the Interaction between the HasR Outer Membrane Receptor and Its Specific TonB-like Protein, HasB

Structural and molecular determinants for the interaction of ExbB from Serratia marcescens and HasB, a TonB paralog

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