Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius

Lena Blagova; Alex Henrique Miller; Megan Bennett; Rosalind Booth; Eleanor J. Dodson; Anne-Kathrin Duhme-Klair; Keith Sanderson Wilson

doi:10.1107/S2059798323004473

Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius

Lena Blagova, Alex Henrique Miller, Megan Bennett, Rosalind Booth, Eleanor J. Dodson, Anne-Kathrin Duhme-Klair^*, Keith Sanderson Wilson^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

Siderophore-binding proteins from two thermophilic bacteria, Geobacillus
stearothermophilus and Parageobacillus thermoglucosidasius, were identified
from a search of sequence databases, cloned and overexpressed. They are
homologues of the well characterized protein CjCeuE from Campylobacter
jejuni. The iron-binding histidine and tyrosine residues are conserved in both
thermophiles. Crystal structures were determined of the apo proteins and of
their complexes with iron(III)-azotochelin and its analogue iron(III)-5-LICAM.
The thermostability of both homologues was shown to be about 20C higher
than that of CjCeuE. Similarly, the tolerance of the homologues to the organic
solvent dimethylformamide (DMF) was enhanced, as reflected by the respective
binding constants for these ligands measured in aqueous buffer at pH 7.5 in the
absence and presence of 10% and 20% DMF. Consequently, these thermophilic
homologues offer advantages in the development of artificial metalloenzymes
using the CeuE family.

Original language	English
Pages (from-to)	694-705
Number of pages	12
Journal	Acta Crystallographica Section D: Structural Biology
Volume	D79
Early online date	11 Jul 2023
DOIs	https://doi.org/10.1107/S2059798323004473
Publication status	Published - 28 Jul 2023

Keywords

thermophilic proteins; siderophore binding; structure; biophysical characterization

Access to Document

10.1107/S2059798323004473Licence: CC BY

Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasiusFinal published version, 1.75 MBLicence: CC BY

1 Conference

Faraday Discussion Meeting Harnessing Non Covalent Interactions
Anne-Kathrin Duhme-Klair (Committee member)
12 Apr 2023 → 14 Apr 2023
Activity: Participating in or organising an event › Conference

2 Finished

21ENGBIO In Cell Assembly of Artificial Imine Reductases for Applications in Whole-Cell Catalysis
Duhme-Klair, A., Grogan, G. J., O'Toole, P. J. & Wilson, K. S.
BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL)
15/02/23 → 14/02/24
Project: Research project (funded) › Research
Redox-reversible artificial metalloenzymes
Duhme-Klair, A., Johnson, S. D. & Wilson, K. S.
27/01/20 → 31/10/23
Project: Research project (funded) › Research

Dataset associated with publication: Kinetic and structural analysis of redox-reversible artificial imine reductases
Miller, A. H. (Data Collector), Martins, I. (Contributor) & Duhme-Klair, A. (Creator), University of York, 29 Jul 2024
DOI: 10.15124/65fe7181-d559-4573-a062-40e064929706
Dataset

Cite this

Blagova, L., Miller, A. H., Bennett, M., Booth, R., Dodson, E. J., Duhme-Klair, A.-K., & Wilson, K. S. (2023). Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius. Acta Crystallographica Section D: Structural Biology, D79, 694-705. https://doi.org/10.1107/S2059798323004473

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title = "Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius",

abstract = "Siderophore-binding proteins from two thermophilic bacteria, Geobacillusstearothermophilus and Parageobacillus thermoglucosidasius, were identifiedfrom a search of sequence databases, cloned and overexpressed. They arehomologues of the well characterized protein CjCeuE from Campylobacterjejuni. The iron-binding histidine and tyrosine residues are conserved in boththermophiles. Crystal structures were determined of the apo proteins and oftheir complexes with iron(III)-azotochelin and its analogue iron(III)-5-LICAM.The thermostability of both homologues was shown to be about 20C higherthan that of CjCeuE. Similarly, the tolerance of the homologues to the organicsolvent dimethylformamide (DMF) was enhanced, as reflected by the respectivebinding constants for these ligands measured in aqueous buffer at pH 7.5 in theabsence and presence of 10% and 20% DMF. Consequently, these thermophilichomologues offer advantages in the development of artificial metalloenzymesusing the CeuE family.",

keywords = "thermophilic proteins; siderophore binding; structure; biophysical characterization",

author = "Lena Blagova and Miller, {Alex Henrique} and Megan Bennett and Rosalind Booth and Dodson, {Eleanor J.} and Anne-Kathrin Duhme-Klair and Wilson, {Keith Sanderson}",

year = "2023",

month = jul,

day = "28",

doi = "10.1107/S2059798323004473",

language = "English",

volume = "D79",

pages = "694--705",

journal = "Acta Crystallographica Section D: Structural Biology",

issn = "2059-7983",

publisher = "Wiley-Blackwell",

}

Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius. / Blagova, Lena ; Miller, Alex Henrique ; Bennett, Megan et al.
In: Acta Crystallographica Section D: Structural Biology, Vol. D79, 28.07.2023, p. 694-705.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius

AU - Blagova, Lena

AU - Miller, Alex Henrique

AU - Bennett, Megan

AU - Booth, Rosalind

AU - Dodson, Eleanor J.

AU - Duhme-Klair, Anne-Kathrin

AU - Wilson, Keith Sanderson

PY - 2023/7/28

Y1 - 2023/7/28

N2 - Siderophore-binding proteins from two thermophilic bacteria, Geobacillusstearothermophilus and Parageobacillus thermoglucosidasius, were identifiedfrom a search of sequence databases, cloned and overexpressed. They arehomologues of the well characterized protein CjCeuE from Campylobacterjejuni. The iron-binding histidine and tyrosine residues are conserved in boththermophiles. Crystal structures were determined of the apo proteins and oftheir complexes with iron(III)-azotochelin and its analogue iron(III)-5-LICAM.The thermostability of both homologues was shown to be about 20C higherthan that of CjCeuE. Similarly, the tolerance of the homologues to the organicsolvent dimethylformamide (DMF) was enhanced, as reflected by the respectivebinding constants for these ligands measured in aqueous buffer at pH 7.5 in theabsence and presence of 10% and 20% DMF. Consequently, these thermophilichomologues offer advantages in the development of artificial metalloenzymesusing the CeuE family.

AB - Siderophore-binding proteins from two thermophilic bacteria, Geobacillusstearothermophilus and Parageobacillus thermoglucosidasius, were identifiedfrom a search of sequence databases, cloned and overexpressed. They arehomologues of the well characterized protein CjCeuE from Campylobacterjejuni. The iron-binding histidine and tyrosine residues are conserved in boththermophiles. Crystal structures were determined of the apo proteins and oftheir complexes with iron(III)-azotochelin and its analogue iron(III)-5-LICAM.The thermostability of both homologues was shown to be about 20C higherthan that of CjCeuE. Similarly, the tolerance of the homologues to the organicsolvent dimethylformamide (DMF) was enhanced, as reflected by the respectivebinding constants for these ligands measured in aqueous buffer at pH 7.5 in theabsence and presence of 10% and 20% DMF. Consequently, these thermophilichomologues offer advantages in the development of artificial metalloenzymesusing the CeuE family.

KW - thermophilic proteins; siderophore binding; structure; biophysical characterization

U2 - 10.1107/S2059798323004473

DO - 10.1107/S2059798323004473

M3 - Article

SN - 2059-7983

VL - D79

SP - 694

EP - 705

JO - Acta Crystallographica Section D: Structural Biology

JF - Acta Crystallographica Section D: Structural Biology

ER -

Thermostable homologues of the periplasmic siderophore-binding protein CeuE from Geobacillus stearothermophilus and Parageobacillus thermoglucosidasius

Abstract

Keywords

Access to Document

Activities

Faraday Discussion Meeting Harnessing Non Covalent Interactions

Projects

21ENGBIO In Cell Assembly of Artificial Imine Reductases for Applications in Whole-Cell Catalysis

Redox-reversible artificial metalloenzymes

Datasets

Dataset associated with publication: Kinetic and structural analysis of redox-reversible artificial imine reductases

Cite this