Tailorable Tetrahelical Bundles as a Toolkit for Redox Studies

Solomon, Lee A.; Witten, Joshua; Kodali, Goutham; Moser, Christopher C.; Dutton, P. Leslie

doi:10.1021/acs.jpcb.2c05119

Title: Tailorable Tetrahelical Bundles as a Toolkit for Redox Studies

Journal Article · Tue Oct 11 00:00:00 EDT 2022 · Journal of Physical Chemistry. B

DOI:https://doi.org/10.1021/acs.jpcb.2c05119· OSTI ID:1891664

^[1]; Witten, Joshua ^[2];

^[3];

^[3]

Department of Chemistry and Biochemistry, George Mason University, Fairfax, Virginia22030, United States
Department of Biology, George Mason University, Fairfax, Virginia22030, United States
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, Pennsylvania19104, United States

Oxidoreductases have evolved over millions of years to perform a variety of metabolic tasks crucial for life. Understanding how these tasks are engineered relies on delivering external electron donors or acceptors to initiate electron transfer reactions. This is a challenge. Small-molecule redox reagents can act indiscriminately, poisoning the cell. Natural redox proteins are more selective, but finding the right partner can be difficult due to the limited number of redox potentials and difficulty tuning them. De novo proteins offer an alternative path. They are robust and can withstand mutations that allow for tailorable changes. They are also devoid of evolutionary artifacts and readily bind redox cofactors. However, no reliable set of engineering principles have been developed that allow for these proteins to be finetuned so their redox midpoint potential (E_m) can form donor/acceptor pairs with any natural oxidoreductase. This work dissects protein-cofactor interactions that can be tuned to modulate redox potentials of acceptors and donors using a mutable de novo designed tetrahelical protein platform with iron tetrapyrrole cofactors as a test case. We show a series of engineered heme b-binding de novo proteins and quantify their resulting effect on E_m. By focusing on the surface charge and buried charges, as well as cofactor placement, chemical modification, and ligation of cofactors, we are able to achieve a broad range of E_m values spanning a range of 330 mV. We anticipate this work will guide the design of proteinaceous tools that can interface with natural oxidoreductases inside and outside the cell while shedding light on how natural proteins modulate E_m values of bound cofactors.

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Research Organization:: George Mason Univ., Fairfax, VA (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences & Engineering Division

Grant/Contract Number:: FG02- 05ER46223; FG02-05ER46223

OSTI ID:: 1891664

Alternate ID(s):: OSTI ID: 1894107

Journal Information:: Journal of Physical Chemistry. B, Journal Name: Journal of Physical Chemistry. B Vol. 126 Journal Issue: 41; ISSN 1520-6106

Publisher:: American Chemical SocietyCopyright Statement

Country of Publication:: United States

Language:: English

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Title: Tailorable Tetrahelical Bundles as a Toolkit for Redox Studies

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