A Versatile Method for Site-Specific Chemical Installation of Aromatic Posttranslational Modification Analogs into Proteins DOI Creative Commons

Xiaoxi Lin,

Shaswati Mandal, Raj V. Nithun

и другие.

Опубликована: Июнь 17, 2024

Posttranslational modifications (PTMs) of proteins play central roles in regulating protein structure, interactome, and functions. A notable modification site is the aromatic side chain Tyr, which undergoes modifications, such as phosphorylation nitration. Despite biological physiological importance Tyr-PTMs, our current understanding mechanisms by these contribute to human health disease remains incomplete. This knowledge gap arises from absence natural amino acids that can mimic PTMs lack synthetic tools for site-specific introduction into proteins. Herein, we describe a facile method chemical installation through palladium-mediate S-C(sp2) bond formation under ambient conditions. We demonstrate incorporation novel Tyr-nitration analogs recombinantly expressed Cys-containing peptides within minutes good yield. To versatility approach, employed it prepare 10 site-specifically modified proteins, including nitrated Myc Max Furthermore, prepared focused library phosphorylated α-Syn protein, enabled first time deciphering role competing conformation aggregation vitro. Our strategy offers advantages over or semi-synthetic approaches, enables rapid selective transfer rarely explored recombinant thus facilitating generation libraries homogeneous post-translationally biomarkers discovery, mechanistic studies, drug discovery.

Язык: Английский

Examining the Role of Threonine Phosphorylation in Ubiquitin’s Function Using Chemical Protein Synthesis DOI Creative Commons

B J Wang,

Chuntong Li, Feng Zhao

и другие.

JACS Au, Год журнала: 2025, Номер unknown

Опубликована: Апрель 21, 2025

The phosphorylation of ubiquitin significantly enhances the complexity code. However, molecular consequences at threonine residues remain largely uncharacterized. In this study, we present an effective method for total chemical synthesis threonine-phosphorylated ubiquitin, producing tens milligrams all six in vivo-identified analogues: pUbT7, pUbT12, pUbT14, pUbT22, pUbT55, and pUbT66. biochemical activities phosphorylated analogues were examined vitro. Our results show that has a differential impact on E2 charging, with residue Thr7 exhibiting significant inhibition. addition, affects E1-E2-E3-mediated assembly deubiquitinase-mediated disassembly polyubiquitin chains site-specific manner. Collectively, work provides new insights into effect

Язык: Английский

Процитировано

1
A Versatile Method for Site-Specific Chemical Installation of Aromatic Posttranslational Modification Analogs into Proteins DOI Creative Commons

Xiaoxi Lin,

Shaswati Mandal, Raj V. Nithun

и другие.

Journal of the American Chemical Society, Год журнала: 2024, Номер 146(37), С. 25788 - 25798

Опубликована: Сен. 3, 2024

Posttranslational modifications (PTMs) of proteins play central roles in regulating the protein structure, interactome, and functions. A notable modification site is aromatic side chain Tyr, which undergoes such as phosphorylation nitration. Despite biological physiological importance Tyr-PTMs, our current understanding mechanisms by these contribute to human health disease remains incomplete. This knowledge gap arises from absence natural amino acids that can mimic PTMs lack synthetic tools for site-specific introduction into proteins. Herein, we describe a facile method chemical installation through palladium-mediated S-C(sp

Язык: Английский

Процитировано

6
A Versatile Method for Site-Specific Chemical Installation of Aromatic Posttranslational Modification Analogs into Proteins DOI Creative Commons

Xiaoxi Lin,

Shaswati Mandal, Raj V. Nithun

и другие.

Опубликована: Июнь 17, 2024

Posttranslational modifications (PTMs) of proteins play central roles in regulating protein structure, interactome, and functions. A notable modification site is the aromatic side chain Tyr, which undergoes modifications, such as phosphorylation nitration. Despite biological physiological importance Tyr-PTMs, our current understanding mechanisms by these contribute to human health disease remains incomplete. This knowledge gap arises from absence natural amino acids that can mimic PTMs lack synthetic tools for site-specific introduction into proteins. Herein, we describe a facile method chemical installation through palladium-mediate S-C(sp2) bond formation under ambient conditions. We demonstrate incorporation novel Tyr-nitration analogs recombinantly expressed Cys-containing peptides within minutes good yield. To versatility approach, employed it prepare 10 site-specifically modified proteins, including nitrated Myc Max Furthermore, prepared focused library phosphorylated α-Syn protein, enabled first time deciphering role competing conformation aggregation vitro. Our strategy offers advantages over or semi-synthetic approaches, enables rapid selective transfer rarely explored recombinant thus facilitating generation libraries homogeneous post-translationally biomarkers discovery, mechanistic studies, drug discovery.

Язык: Английский

Процитировано

1