A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation

Abstract Natural enzymes use local environments to tune the reactivity of amino acid side chains. In searching for small peptides with similar properties, we discovered a four-residue π-clamp motif (Phe-Cys-Pro-Phe) for regio- and chemoselective arylation of cysteine in ribosomally produced proteins...

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Autores principales:	Peng Dai, Jonathan K. Williams, Chi Zhang, Matthew Welborn, James J. Shepherd, Tianyu Zhu, Troy Van Voorhis, Mei Hong, Bradley L. Pentelute
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/65ff2ecec9fc40949a8979455dbaebf8
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spelling	oai:doaj.org-article:65ff2ecec9fc40949a8979455dbaebf82021-12-02T11:40:31ZA structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation10.1038/s41598-017-08402-22045-2322https://doaj.org/article/65ff2ecec9fc40949a8979455dbaebf82017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08402-2https://doaj.org/toc/2045-2322Abstract Natural enzymes use local environments to tune the reactivity of amino acid side chains. In searching for small peptides with similar properties, we discovered a four-residue π-clamp motif (Phe-Cys-Pro-Phe) for regio- and chemoselective arylation of cysteine in ribosomally produced proteins. Here we report mutational, computational, and structural findings directed toward elucidating the molecular factors that drive π-clamp-mediated arylation. We show the significance of a trans conformation prolyl amide bond for the π-clamp reactivity. The π-clamp cysteine arylation reaction enthalpy of activation (ΔH‡) is significantly lower than a non-π-clamp cysteine. Solid-state NMR chemical shifts indicate the prolyl amide bond in the π-clamp motif adopts a 1:1 ratio of the cis and trans conformation, while in the reaction product Pro3 was exclusively in trans. In two structural models of the perfluoroarylated product, distinct interactions at 4.7 Å between Phe1 side chain and perfluoroaryl electrophile moiety are observed. Further, solution 19F NMR and isothermal titration calorimetry measurements suggest interactions between hydrophobic side chains in a π-clamp mutant and the perfluoroaryl probe. These studies led us to design a π-clamp mutant with an 85-fold rate enhancement. These findings will guide us toward the discovery of small reactive peptides to facilitate abiotic chemistry in water.Peng DaiJonathan K. WilliamsChi ZhangMatthew WelbornJames J. ShepherdTianyu ZhuTroy Van VoorhisMei HongBradley L. PenteluteNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Peng Dai Jonathan K. Williams Chi Zhang Matthew Welborn James J. Shepherd Tianyu Zhu Troy Van Voorhis Mei Hong Bradley L. Pentelute A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
description	Abstract Natural enzymes use local environments to tune the reactivity of amino acid side chains. In searching for small peptides with similar properties, we discovered a four-residue π-clamp motif (Phe-Cys-Pro-Phe) for regio- and chemoselective arylation of cysteine in ribosomally produced proteins. Here we report mutational, computational, and structural findings directed toward elucidating the molecular factors that drive π-clamp-mediated arylation. We show the significance of a trans conformation prolyl amide bond for the π-clamp reactivity. The π-clamp cysteine arylation reaction enthalpy of activation (ΔH‡) is significantly lower than a non-π-clamp cysteine. Solid-state NMR chemical shifts indicate the prolyl amide bond in the π-clamp motif adopts a 1:1 ratio of the cis and trans conformation, while in the reaction product Pro3 was exclusively in trans. In two structural models of the perfluoroarylated product, distinct interactions at 4.7 Å between Phe1 side chain and perfluoroaryl electrophile moiety are observed. Further, solution 19F NMR and isothermal titration calorimetry measurements suggest interactions between hydrophobic side chains in a π-clamp mutant and the perfluoroaryl probe. These studies led us to design a π-clamp mutant with an 85-fold rate enhancement. These findings will guide us toward the discovery of small reactive peptides to facilitate abiotic chemistry in water.
format	article
author	Peng Dai Jonathan K. Williams Chi Zhang Matthew Welborn James J. Shepherd Tianyu Zhu Troy Van Voorhis Mei Hong Bradley L. Pentelute
author_facet	Peng Dai Jonathan K. Williams Chi Zhang Matthew Welborn James J. Shepherd Tianyu Zhu Troy Van Voorhis Mei Hong Bradley L. Pentelute
author_sort	Peng Dai
title	A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
title_short	A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
title_full	A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
title_fullStr	A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
title_full_unstemmed	A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
title_sort	structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/65ff2ecec9fc40949a8979455dbaebf8
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A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation

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