Biochemical and Structural Characterization of XoxG and XoxJ and Their Roles in Lanthanide-Dependent Methanol Dehydrogenase Activity

Emily R. Featherston, Hannah R. Rose, Molly J. McBride, Ellison M. Taylor, Amie Kathleen Boal, Joseph Alfred Cotruvo, Jr.

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Lanthanide (Ln)-dependent methanol dehydrogenases (MDHs) have recently been shown to be widespread in methylotrophic bacteria. Along with the core MDH protein, XoxF, these systems contain two other proteins, XoxG (a c-type cytochrome) and XoxJ (a periplasmic binding protein of unknown function), about which little is known. In this work, we have biochemically and structurally characterized these proteins from the methyltroph Methylobacterium extorquens AM1. In contrast to results obtained in an artificial assay system, assays of XoxFs metallated with LaIII, CeIII, and NdIII using their physiological electron acceptor, XoxG, display Ln-independent activities, but the Km for XoxG markedly increases from La to Nd. This result suggests that XoxG′s redox properties are tuned specifically for lighter Lns in XoxF, an interpretation supported by the unusually low reduction potential of XoxG (+172 mV). The X-ray crystal structure of XoxG provides a structural basis for this reduction potential and insight into the XoxG–XoxF interaction. Finally, the X-ray crystal structure of XoxJ reveals a large hydrophobic cleft and suggests a role in the activation of XoxF. These studies enrich our understanding of the underlying chemical principles that enable the activity of XoxF with multiple lanthanides in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)2360-2372
Number of pages13
JournalChemBioChem
Volume20
Issue number18
DOIs
StatePublished - Sep 16 2019

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Lanthanoid Series Elements
Assays
Methylobacterium extorquens
Crystal structure
Periplasmic Binding Proteins
X-Rays
Cytochrome c Group
X rays
Proteins
Oxidation-Reduction
Bacteria
Chemical activation
Electrons
alcohol dehydrogenase (acceptor)

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

Cite this

Featherston, Emily R. ; Rose, Hannah R. ; McBride, Molly J. ; Taylor, Ellison M. ; Boal, Amie Kathleen ; Cotruvo, Jr., Joseph Alfred. / Biochemical and Structural Characterization of XoxG and XoxJ and Their Roles in Lanthanide-Dependent Methanol Dehydrogenase Activity. In: ChemBioChem. 2019 ; Vol. 20, No. 18. pp. 2360-2372.
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Biochemical and Structural Characterization of XoxG and XoxJ and Their Roles in Lanthanide-Dependent Methanol Dehydrogenase Activity. / Featherston, Emily R.; Rose, Hannah R.; McBride, Molly J.; Taylor, Ellison M.; Boal, Amie Kathleen; Cotruvo, Jr., Joseph Alfred.

In: ChemBioChem, Vol. 20, No. 18, 16.09.2019, p. 2360-2372.

Research output: Contribution to journalArticle

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T1 - Biochemical and Structural Characterization of XoxG and XoxJ and Their Roles in Lanthanide-Dependent Methanol Dehydrogenase Activity

AU - Featherston, Emily R.

AU - Rose, Hannah R.

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AU - Boal, Amie Kathleen

AU - Cotruvo, Jr., Joseph Alfred

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AB - Lanthanide (Ln)-dependent methanol dehydrogenases (MDHs) have recently been shown to be widespread in methylotrophic bacteria. Along with the core MDH protein, XoxF, these systems contain two other proteins, XoxG (a c-type cytochrome) and XoxJ (a periplasmic binding protein of unknown function), about which little is known. In this work, we have biochemically and structurally characterized these proteins from the methyltroph Methylobacterium extorquens AM1. In contrast to results obtained in an artificial assay system, assays of XoxFs metallated with LaIII, CeIII, and NdIII using their physiological electron acceptor, XoxG, display Ln-independent activities, but the Km for XoxG markedly increases from La to Nd. This result suggests that XoxG′s redox properties are tuned specifically for lighter Lns in XoxF, an interpretation supported by the unusually low reduction potential of XoxG (+172 mV). The X-ray crystal structure of XoxG provides a structural basis for this reduction potential and insight into the XoxG–XoxF interaction. Finally, the X-ray crystal structure of XoxJ reveals a large hydrophobic cleft and suggests a role in the activation of XoxF. These studies enrich our understanding of the underlying chemical principles that enable the activity of XoxF with multiple lanthanides in vitro and in vivo.

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