Dataset Information


Formal Reduction Potentials of Difluorotyrosine and Trifluorotyrosine Protein Residues: Defining the Thermodynamics of Multistep Radical Transfer.

ABSTRACT: Redox-active tyrosines (Ys) play essential roles in enzymes involved in primary metabolism including energy transduction and deoxynucleotide production catalyzed by ribonucleotide reductases (RNRs). Thermodynamic characterization of Ys in solution and in proteins remains a challenge due to the high reduction potentials involved and the reactive nature of the radical state. The structurally characterized ?3Y model protein has allowed the first determination of formal reduction potentials (E°') for a Y residing within a protein (Berry, B. W.; Mart??nez-Rivera, M. C.; Tommos, C. Proc. Natl. Acad. Sci. U. S. A. 2012, 109, 9739-9743). Using Schultz's technology, a series of fluorotyrosines (FnY, n = 2 or 3) was site-specifically incorporated into ?3Y. The global protein properties of the resulting ?3(3,5)F2Y, ?3(2,3,5)F3Y, ?3(2,3)F2Y and ?3(2,3,6)F3Y variants are essentially identical to those of ?3Y. A protein film square-wave voltammetry approach was developed to successfully obtain reversible voltammograms and E°'s of the very high-potential ?3FnY proteins. E°'(pH 5.5; ?3FnY(O•/OH)) spans a range of 1040 ± 3 mV to 1200 ± 3 mV versus the normal hydrogen electrode. This is comparable to the potentials of the most oxidizing redox cofactors in nature. The FnY analogues, and the ability to site-specifically incorporate them into any protein of interest, provide new tools for mechanistic studies on redox-active Ys in proteins and on functional and aberrant hole-transfer reactions in metallo-enzymes. The former application is illustrated here by using the determined ?3FnY ?E°'s to model the thermodynamics of radical-transfer reactions in FnY-RNRs and to experimentally test and support the key prediction made.

SUBMITTER: Ravichandran KR 

PROVIDER: S-EPMC5651514 | BioStudies | 2017-01-01

REPOSITORIES: biostudies

Similar Datasets

2013-01-01 | S-EPMC4076196 | BioStudies
2016-01-01 | S-EPMC4929525 | BioStudies
2016-01-01 | S-EPMC5224885 | BioStudies
2011-01-01 | S-EPMC3188361 | BioStudies
2018-01-01 | S-EPMC5762255 | BioStudies
2013-01-01 | S-EPMC3823682 | BioStudies
2018-01-01 | S-EPMC5985984 | BioStudies
2017-01-01 | S-EPMC5689411 | BioStudies
2008-01-01 | S-EPMC2567162 | BioStudies
2006-01-01 | S-EPMC2532977 | BioStudies