The biophysical basis of protein domain compatibility

Coyote‐Maestas, Willow; Nedrud, David; Suma, Antonio; He, Yungui; Matreyek, Kenneth A.; Fowler, Douglas M.; Carnevale, Vincenzo; Myers, Chad L.; Schmidt, Daniel

doi:10.1101/2020.12.09.418442

Cited by 2 publications

(4 citation statements)

References 58 publications

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“…Cα B factors were obtained from the crystal structure for each residue in MtrA using the Biopython Bio.PDB package , and averaged for the residue preceding and the residue following each insertion site. Per-residue stiffness estimates were obtained from a Cα anisotropic network model of the entire MtrCAB complex (without hemes) using the ProDy suite, , based on previously described protocols. , …”

Section: Methodsmentioning

confidence: 99%

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Determinants of Multiheme Cytochrome Extracellular Electron Transfer Uncovered by Systematic Peptide Insertion

et al. 2022

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The multiheme cytochrome MtrA enables microbial respiration by transferring electrons across the outer membrane to extracellular electron acceptors. While structural studies have identified residues that mediate the binding of MtrA to hemes and to other cytochromes that facilitate extracellular electron transfer (EET), the relative importance of these interactions for EET is not known. To better understand EET, we evaluated how insertion of an octapeptide across all MtrA backbone locations affects Shewanella oneidensis MR-1 respiration on Fe(III). The EET efficiency was found to be inversely correlated with the proximity of the insertion to the heme prosthetic groups. Mutants with decreased EET efficiencies also arose from insertions in a subset of the regions that make residue–residue contacts with the porin MtrB, while all sites contacting the extracellular cytochrome MtrC presented high peptide insertion tolerance. MtrA variants having peptide insertions within the CXXCH motifs that coordinate heme cofactors retained some ability to support respiration on Fe(III), although these variants presented significantly decreased EET efficiencies. Furthermore, the fitness of cells expressing different MtrA variants under Fe(III) respiration conditions correlated with anode reduction. The peptide insertion profile, which represents the first comprehensive sequence–structure–function map for a multiheme cytochrome, implicates MtrA as a strategic protein engineering target for the regulation of EET.

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Section: Methodsmentioning

confidence: 99%

“…Per-residue stiffness estimates were obtained from a Cα anisotropic network model of the entire MtrCAB complex (without hemes) using the ProDy suite, 47,48 based on previously described protocols. 44,49 Statistics. All error bars represent ±1 standard deviation calculated from three or more biological replicates.…”

Section: ■ Methodsmentioning

confidence: 99%

Determinants of Multiheme Cytochrome Extracellular Electron Transfer Uncovered by Systematic Peptide Insertion

et al. 2022

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

“…Per-residue stiffness estimates were obtained from a Cα anisotropic network model of the entire MtrCAB complex (without hemes) using the ProDy suite, 71,72 based on previously described protocols. 68,73 Statistics. All error bars represent ±1 standard deviation calculated from three or more biological replicates.…”

Section: Moo3 Nanoparticle Assaymentioning

confidence: 99%

Determinants of multiheme cytochrome extracellular electron transfer uncovered by systematic peptide insertion

Campbell

Atkinson

Carpenter

et al. 2022

Preprint

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The multiheme cytochrome MtrA enables microbial respiration by transferring electrons across the outer membrane to extracellular electron acceptors. While structural studies have identified residues that mediate MtrA binding to hemes and to other cytochromes that facilitate extracellular electron transfer (EET), the relative importance of these interactions for EET is not known. To better understand EET, we evaluated how insertion of an octapeptide across all MtrA backbone locations affects Shewanella oneidensis MR-1 respiration on Fe(III). EET efficiency was found to be inversely correlated with insertion proximity to the heme prosthetic groups. Mutants with decreased EET also arose from insertions in a subset of the regions that make residue-residue contacts with the porin MtrB, while all sites contacting the extracellular MtrC presented high peptide insertion tolerance. MtrA variants having peptide insertions within the CXXCH motifs that coordinate heme cofactors retained some ability to support respiration on Fe(III), although these variants presented significantly decreased EET. Furthermore, the fitness of cells expressing different MtrA variants under Fe(III)-respiring conditions correlated with anode reduction. The peptide-insertion profile, which represents the first comprehensive sequence-structure-function map for a multiheme cytochrome, implicates MtrA as a strategic protein engineering target for regulating EET.

show abstract

The biophysical basis of protein domain compatibility

Cited by 2 publications

References 58 publications

Determinants of Multiheme Cytochrome Extracellular Electron Transfer Uncovered by Systematic Peptide Insertion

Determinants of Multiheme Cytochrome Extracellular Electron Transfer Uncovered by Systematic Peptide Insertion

Determinants of multiheme cytochrome extracellular electron transfer uncovered by systematic peptide insertion

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