Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization

Succination is a non-enzymatic post-translational modification of cysteine (Cys) residues, resulting in the formation of <i>S</i>-(2-succino)cysteine (2SC). While hundreds of 2SC-modified proteins have been identified and are associated with the dysfunction of proteins, the underlying mo...

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Main Authors: Hiroshi Yamaguchi, Hikari Sugawa, Himeno Takahashi, Ryoji Nagai
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/3/571
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author Hiroshi Yamaguchi
Hikari Sugawa
Himeno Takahashi
Ryoji Nagai
author_facet Hiroshi Yamaguchi
Hikari Sugawa
Himeno Takahashi
Ryoji Nagai
author_sort Hiroshi Yamaguchi
collection DOAJ
description Succination is a non-enzymatic post-translational modification of cysteine (Cys) residues, resulting in the formation of <i>S</i>-(2-succino)cysteine (2SC). While hundreds of 2SC-modified proteins have been identified and are associated with the dysfunction of proteins, the underlying molecular mechanisms remain poorly understood. Conventional methods for synthesizing succinated compounds, such as 2SC, often require prolonged reaction times and/or HCl hydrolysis. In this study, we present a rapid and efficient synthesis method for succinated compounds using maleic anhydride, enabling more effective in vivo studies of succination mechanisms. This method was tested on thiol compounds with varying molecular weights, including Cys derivatives, Cys-containing peptides, and reduced bovine serum albumin. By incubating these compounds in an aqueous buffer with maleic anhydride dissolved in an organic solvent like diethyl ether, we achieved significantly improved succination efficiency compared to conventional methods. The succination efficiency using maleic anhydride surpassed that of fumaric acid or maleic acid. Notably, this approach facilitated the succination of amino acids, peptides, and proteins within minutes at 25 °C, without requiring acid hydrolysis. Our findings provide a straightforward, time-efficient strategy for synthesizing succinated thiol compounds, offering a valuable tool to enhance the understanding of succination’s molecular mechanisms and its role in protein function and dysfunction.
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spelling doaj-art-8fd37413b1fa4b4f9ff2c683f9beee6f2025-08-20T03:12:35ZengMDPI AGMolecules1420-30492025-01-0130357110.3390/molecules30030571Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride DerivatizationHiroshi Yamaguchi0Hikari Sugawa1Himeno Takahashi2Ryoji Nagai3Department of Food and Life Science, School of Agriculture, Tokai University, 871-12 Sugido, Mashiki, Kamimashiki, Kumamoto 861-2205, JapanDepartment of Food and Life Science, School of Agriculture, Tokai University, 871-12 Sugido, Mashiki, Kamimashiki, Kumamoto 861-2205, JapanGraduate School of Bioscience, Tokai University, 871-12 Sugido, Mashiki, Kamimashiki, Kumamoto 861-2205, JapanDepartment of Food and Life Science, School of Agriculture, Tokai University, 871-12 Sugido, Mashiki, Kamimashiki, Kumamoto 861-2205, JapanSuccination is a non-enzymatic post-translational modification of cysteine (Cys) residues, resulting in the formation of <i>S</i>-(2-succino)cysteine (2SC). While hundreds of 2SC-modified proteins have been identified and are associated with the dysfunction of proteins, the underlying molecular mechanisms remain poorly understood. Conventional methods for synthesizing succinated compounds, such as 2SC, often require prolonged reaction times and/or HCl hydrolysis. In this study, we present a rapid and efficient synthesis method for succinated compounds using maleic anhydride, enabling more effective in vivo studies of succination mechanisms. This method was tested on thiol compounds with varying molecular weights, including Cys derivatives, Cys-containing peptides, and reduced bovine serum albumin. By incubating these compounds in an aqueous buffer with maleic anhydride dissolved in an organic solvent like diethyl ether, we achieved significantly improved succination efficiency compared to conventional methods. The succination efficiency using maleic anhydride surpassed that of fumaric acid or maleic acid. Notably, this approach facilitated the succination of amino acids, peptides, and proteins within minutes at 25 °C, without requiring acid hydrolysis. Our findings provide a straightforward, time-efficient strategy for synthesizing succinated thiol compounds, offering a valuable tool to enhance the understanding of succination’s molecular mechanisms and its role in protein function and dysfunction.https://www.mdpi.com/1420-3049/30/3/571succinationmaleic anhydridethiol compoundMichael addition reaction
spellingShingle Hiroshi Yamaguchi
Hikari Sugawa
Himeno Takahashi
Ryoji Nagai
Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization
Molecules
succination
maleic anhydride
thiol compound
Michael addition reaction
title Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization
title_full Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization
title_fullStr Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization
title_full_unstemmed Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization
title_short Rapid and Efficient Synthesis of Succinated Thiol Compounds via Maleic Anhydride Derivatization
title_sort rapid and efficient synthesis of succinated thiol compounds via maleic anhydride derivatization
topic succination
maleic anhydride
thiol compound
Michael addition reaction
url https://www.mdpi.com/1420-3049/30/3/571
work_keys_str_mv AT hiroshiyamaguchi rapidandefficientsynthesisofsuccinatedthiolcompoundsviamaleicanhydridederivatization
AT hikarisugawa rapidandefficientsynthesisofsuccinatedthiolcompoundsviamaleicanhydridederivatization
AT himenotakahashi rapidandefficientsynthesisofsuccinatedthiolcompoundsviamaleicanhydridederivatization
AT ryojinagai rapidandefficientsynthesisofsuccinatedthiolcompoundsviamaleicanhydridederivatization