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Defining the Role of Tyrosine and Rational Tuning of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine Analogs
[Image: see text] While a conserved tyrosine (Tyr) is found in oxidases, the roles of phenol ring pK(a) and reduction potential in O(2) reduction have not been defined despite many years of research on numerous oxidases and their models. These issues represent major challenges in our understanding o...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2015
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4676419/ https://www.ncbi.nlm.nih.gov/pubmed/25672571 http://dx.doi.org/10.1021/ja5109936 |
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author | Yu, Yang Lv, Xiaoxuan Li, Jiasong Zhou, Qing Cui, Chang Hosseinzadeh, Parisa Mukherjee, Arnab Nilges, Mark J. Wang, Jiangyun Lu, Yi |
author_facet | Yu, Yang Lv, Xiaoxuan Li, Jiasong Zhou, Qing Cui, Chang Hosseinzadeh, Parisa Mukherjee, Arnab Nilges, Mark J. Wang, Jiangyun Lu, Yi |
author_sort | Yu, Yang |
collection | PubMed |
description | [Image: see text] While a conserved tyrosine (Tyr) is found in oxidases, the roles of phenol ring pK(a) and reduction potential in O(2) reduction have not been defined despite many years of research on numerous oxidases and their models. These issues represent major challenges in our understanding of O(2) reduction mechanism in bioenergetics. Through genetic incorporation of unnatural amino acid analogs of Tyr, with progressively decreasing pK(a) of the phenol ring and increasing reduction potential, in the active site of a functional model of oxidase in myoglobin, a linear dependence of both the O(2) reduction activity and the fraction of H(2)O formation with the pK(a) of the phenol ring has been established. By using these unnatural amino acids as spectroscopic probe, we have provided conclusive evidence for the location of a Tyr radical generated during reaction with H(2)O(2), by the distinctive hyperfine splitting patterns of the halogenated tyrosines and one of its deuterated derivatives incorporated at the 33 position of the protein. These results demonstrate for the first time that enhancing the proton donation ability of the Tyr enhances the oxidase activity, allowing the Tyr analogs to augment enzymatic activity beyond that of natural Tyr. |
format | Online Article Text |
id | pubmed-4676419 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-46764192016-02-11 Defining the Role of Tyrosine and Rational Tuning of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine Analogs Yu, Yang Lv, Xiaoxuan Li, Jiasong Zhou, Qing Cui, Chang Hosseinzadeh, Parisa Mukherjee, Arnab Nilges, Mark J. Wang, Jiangyun Lu, Yi J Am Chem Soc [Image: see text] While a conserved tyrosine (Tyr) is found in oxidases, the roles of phenol ring pK(a) and reduction potential in O(2) reduction have not been defined despite many years of research on numerous oxidases and their models. These issues represent major challenges in our understanding of O(2) reduction mechanism in bioenergetics. Through genetic incorporation of unnatural amino acid analogs of Tyr, with progressively decreasing pK(a) of the phenol ring and increasing reduction potential, in the active site of a functional model of oxidase in myoglobin, a linear dependence of both the O(2) reduction activity and the fraction of H(2)O formation with the pK(a) of the phenol ring has been established. By using these unnatural amino acids as spectroscopic probe, we have provided conclusive evidence for the location of a Tyr radical generated during reaction with H(2)O(2), by the distinctive hyperfine splitting patterns of the halogenated tyrosines and one of its deuterated derivatives incorporated at the 33 position of the protein. These results demonstrate for the first time that enhancing the proton donation ability of the Tyr enhances the oxidase activity, allowing the Tyr analogs to augment enzymatic activity beyond that of natural Tyr. American Chemical Society 2015-02-11 2015-04-15 /pmc/articles/PMC4676419/ /pubmed/25672571 http://dx.doi.org/10.1021/ja5109936 Text en Copyright © 2015 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Yu, Yang Lv, Xiaoxuan Li, Jiasong Zhou, Qing Cui, Chang Hosseinzadeh, Parisa Mukherjee, Arnab Nilges, Mark J. Wang, Jiangyun Lu, Yi Defining the Role of Tyrosine and Rational Tuning of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine Analogs |
title | Defining
the Role of Tyrosine and Rational Tuning
of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine
Analogs |
title_full | Defining
the Role of Tyrosine and Rational Tuning
of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine
Analogs |
title_fullStr | Defining
the Role of Tyrosine and Rational Tuning
of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine
Analogs |
title_full_unstemmed | Defining
the Role of Tyrosine and Rational Tuning
of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine
Analogs |
title_short | Defining
the Role of Tyrosine and Rational Tuning
of Oxidase Activity by Genetic Incorporation of Unnatural Tyrosine
Analogs |
title_sort | defining
the role of tyrosine and rational tuning
of oxidase activity by genetic incorporation of unnatural tyrosine
analogs |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4676419/ https://www.ncbi.nlm.nih.gov/pubmed/25672571 http://dx.doi.org/10.1021/ja5109936 |
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