首页|Tuning reduction potentials of type 1 copper center in azurin by replacing a histidine ligand with its isostructural analogues
Tuning reduction potentials of type 1 copper center in azurin by replacing a histidine ligand with its isostructural analogues
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NSTL
Elsevier
? 2022 Elsevier Inc.Type 1 copper proteins have a conserved ligand set of one cysteine and two histidines, with many proteins, such as azurin, also containing an axial methionine. While the cysteine and methionine in azurin have been replaced with their respective isostructural analogues of unnatural amino acids to reveal their roles in tuning electronic structures and functional properties, such as reduction potentials (E°′), the histidine ligands have not been probed in this way. We herein report the substitution of His117 in azurin with three unnatural isostructural analogues, 5-nitrohistidine(Ntr), thiazolylalanine(SHis) and 1-methylhistidine(MeH) by expressed protein ligation. While UV–vis absorption and electron paramagnetic resonance spectroscopies confirm that isostructural replacement results in minimal structural change in the Cu(II) state, the E°′ of these variants increases with increasing pKa of the δ nitrogens of the imidazole. This counter-intuitive relationship between E°′ of the protein and pKa of the sidechain group suggests additional factors may play a role in tuning E°′.
Blue copperCupredoxinsElectron transferProtein engineeringRedoxUnnatural amino acids
Yu Y.、Lu Y.、Marshall N.M.、Garner D.K.、Nilges M.J.
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Center of Biophysics and Quantitative Biology University of Illinois at Urbana?Champaign
Department of Chemistry University of Illinois at Urbana?Champaign
Illinois EPR Research Center University of Illinois at Urbana?Champaign
NSTL
Elsevier
