Incorporation of a molybdenum atom in a Rubredoxin-type Centre of a de novo-designed α3DIV-L21C three-helical bundle peptide

Pedro M. S. Bragança; Marta S. P. Carepo; Sofia R. Pauleta; Tyler Pinter; Maddalena Elia; Cristina M. Cordas; Isabel Moura; Vicent Pecoraro; José J. G. Moura

doi:10.1016/j.jinorgbio.2022.112096

Incorporation of a molybdenum atom in a Rubredoxin-type Centre of a de novo-designed α3DIV-L21C three-helical bundle peptide

Pedro M. S. Bragança, Marta S. P. Carepo, Sofia R. Pauleta, Tyler Pinter, Maddalena Elia, Cristina M. Cordas, Isabel Moura, Vicent Pecoraro, José J. G. Moura

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Abstract

The rational design and functionalization of small, simple, and stable peptides scaffolds is an attractive avenue to mimic catalytic metal-centres of complex proteins, relevant for the design of metalloenzymes with environmental, biotechnological and health impacts. The de novo designed α3DIV-L21C framework has a rubredoxin-like metal binding site and was used in this work to incorporate a Mo-atom. Thermostability studies using differential scanning calorimetry showed an increase of 4 °C in the melting temperature of the Mo-α3DIV-L21C when compared to the apo-α3DIV-L21C. Circular dichroism in the visible and far-UV regions corroborated these results showing that Mo incorporation provides stability to the peptide even though there were almost no differences observed in the secondary structure. A formal reduction potential of ∼ −408 mV vs. NHE, pH 7.6 was determined. Combining electrochemical results, EPR and UV–visible data we discuss the oxidation state of the molybdenum centre in Mo-α3DIV-L21C and propose that is mainly in a Mo (VI) oxidation state.

Original language	English
Article number	112096
Pages (from-to)	1-8
Number of pages	8
Journal	Journal of Inorganic Biochemistry
Volume	240
Early online date	Dec 2022
DOIs	https://doi.org/10.1016/j.jinorgbio.2022.112096
Publication status	Published - Mar 2023

Keywords

A de novo protein design
Three-helix bundle
Molybdenum
Models of molybdenum-containing enzymes
Rubredoxin
Tetracysteinyl coordination

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Bragança, P. M. S., Carepo, M. S. P., Pauleta, S. R., Pinter, T., Elia, M., Cordas, C. M., Moura, I., Pecoraro, V., & Moura, J. J. G. (2023). Incorporation of a molybdenum atom in a Rubredoxin-type Centre of a de novo-designed α3DIV-L21C three-helical bundle peptide. Journal of Inorganic Biochemistry, 240, 1-8. Article 112096. Advance online publication. https://doi.org/10.1016/j.jinorgbio.2022.112096

@article{3e49a67c8d524be8a39d7b43f16424dd,

title = "Incorporation of a molybdenum atom in a Rubredoxin-type Centre of a de novo-designed α3DIV-L21C three-helical bundle peptide",

abstract = "The rational design and functionalization of small, simple, and stable peptides scaffolds is an attractive avenue to mimic catalytic metal-centres of complex proteins, relevant for the design of metalloenzymes with environmental, biotechnological and health impacts. The de novo designed α3DIV-L21C framework has a rubredoxin-like metal binding site and was used in this work to incorporate a Mo-atom. Thermostability studies using differential scanning calorimetry showed an increase of 4 °C in the melting temperature of the Mo-α3DIV-L21C when compared to the apo-α3DIV-L21C. Circular dichroism in the visible and far-UV regions corroborated these results showing that Mo incorporation provides stability to the peptide even though there were almost no differences observed in the secondary structure. A formal reduction potential of ∼ −408 mV vs. NHE, pH 7.6 was determined. Combining electrochemical results, EPR and UV–visible data we discuss the oxidation state of the molybdenum centre in Mo-α3DIV-L21C and propose that is mainly in a Mo (VI) oxidation state.",

keywords = "A de novo protein design, Three-helix bundle, Molybdenum, Models of molybdenum-containing enzymes, Rubredoxin, Tetracysteinyl coordination",

author = "Bragan{\c c}a, {Pedro M. S.} and Carepo, {Marta S. P.} and Pauleta, {Sofia R.} and Tyler Pinter and Maddalena Elia and Cordas, {Cristina M.} and Isabel Moura and Vicent Pecoraro and Moura, {Jos{\'e} J. G.}",

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year = "2023",

month = mar,

doi = "10.1016/j.jinorgbio.2022.112096",

language = "English",

volume = "240",

pages = "1--8",

journal = "Journal of Inorganic Biochemistry",

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AU - Bragança, Pedro M. S.

AU - Carepo, Marta S. P.

AU - Pauleta, Sofia R.

AU - Pinter, Tyler

AU - Elia, Maddalena

AU - Cordas, Cristina M.

AU - Moura, Isabel

AU - Pecoraro, Vicent

AU - Moura, José J. G.

N1 - info:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBTA-BTA%2F0935%2F2020/PT# info:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBIA-BQM%2F29442%2F2017/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F50006%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F50006%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/LA%2FP%2F0008%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/LA%2FP%2F0140%2F2020/PT# PB would thank the PTNMRPhD (PD/00065/2013). VLP thanks the NIH for support (GM141086).

PY - 2023/3

Y1 - 2023/3

N2 - The rational design and functionalization of small, simple, and stable peptides scaffolds is an attractive avenue to mimic catalytic metal-centres of complex proteins, relevant for the design of metalloenzymes with environmental, biotechnological and health impacts. The de novo designed α3DIV-L21C framework has a rubredoxin-like metal binding site and was used in this work to incorporate a Mo-atom. Thermostability studies using differential scanning calorimetry showed an increase of 4 °C in the melting temperature of the Mo-α3DIV-L21C when compared to the apo-α3DIV-L21C. Circular dichroism in the visible and far-UV regions corroborated these results showing that Mo incorporation provides stability to the peptide even though there were almost no differences observed in the secondary structure. A formal reduction potential of ∼ −408 mV vs. NHE, pH 7.6 was determined. Combining electrochemical results, EPR and UV–visible data we discuss the oxidation state of the molybdenum centre in Mo-α3DIV-L21C and propose that is mainly in a Mo (VI) oxidation state.

AB - The rational design and functionalization of small, simple, and stable peptides scaffolds is an attractive avenue to mimic catalytic metal-centres of complex proteins, relevant for the design of metalloenzymes with environmental, biotechnological and health impacts. The de novo designed α3DIV-L21C framework has a rubredoxin-like metal binding site and was used in this work to incorporate a Mo-atom. Thermostability studies using differential scanning calorimetry showed an increase of 4 °C in the melting temperature of the Mo-α3DIV-L21C when compared to the apo-α3DIV-L21C. Circular dichroism in the visible and far-UV regions corroborated these results showing that Mo incorporation provides stability to the peptide even though there were almost no differences observed in the secondary structure. A formal reduction potential of ∼ −408 mV vs. NHE, pH 7.6 was determined. Combining electrochemical results, EPR and UV–visible data we discuss the oxidation state of the molybdenum centre in Mo-α3DIV-L21C and propose that is mainly in a Mo (VI) oxidation state.

KW - A de novo protein design

KW - Three-helix bundle

KW - Molybdenum

KW - Models of molybdenum-containing enzymes

KW - Rubredoxin

KW - Tetracysteinyl coordination

U2 - 10.1016/j.jinorgbio.2022.112096

DO - 10.1016/j.jinorgbio.2022.112096

M3 - Article

C2 - 36603242

SN - 0162-0134

VL - 240

SP - 1

EP - 8

JO - Journal of Inorganic Biochemistry

JF - Journal of Inorganic Biochemistry

M1 - 112096

ER -

Incorporation of a molybdenum atom in a Rubredoxin-type Centre of a de novo-designed α3DIV-L21C three-helical bundle peptide

Abstract

Keywords

UN SDGs

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