Reverse Proteolysis Promoted by in Situ Generated Peptide Ester Fragments
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- 作者：Nicole Wehofsky ; Norman Koglin ; Sven Thust ; and Frank Bordusa
- 刊名：Journal of the American Chemical Society
- 出版年：2003
- 出版时间：May 21, 2003
- 年：2003
- 卷：125
- 期：20
- 页码：6126 - 6133
- 全文大小：120K
- 年卷期：v.125,no.20(May 21, 2003)
- ISSN：1520-5126
文摘
In this contribution we describe a general synthesis concept for the in situ preparation of proteasespecific reactants using methyl thioesters as universal precursors. The precursor esters are readily availableby standard synthesis procedures and can be used directly as reactants for protease-mediated peptidecoupling reactions. Alternatively, they can serve as initial building blocks for the in situ preparation of varioustypes of substrate mimetics. The synthesis of the latter is achieved by a one-pot spontaneoustransthioesterification reaction of the parent thioester (Y-(Xaa)n-SMe
Y-(Xaa)n-SR; R: CH2CH2COOH,CH2C6H5, C6H4NHC(:NH)NH2), which proceeds efficiently in both a sequential manner and parallel to thesubsequent enzymatic reaction. The resulting substrate mimetics act as efficient acyl donor componentsand show the typical behavior of substrate mimicry enabling irreversible reactions with originally nonspecificacyl moieties. Neither a workup of the substrate mimetic intermediate nor changes of the reaction conditionsduring the whole synthesis process are required. Model peptide syntheses using trypsin, 
-chymotrypsin,and V8 protease as the biocatalysts proved the function of the approach and illustrated its synthetic valuefor protease-mediated reactions and the compatibility of the approach with state-of-the-art solid-phasepeptide ester synthesis methods.
Y-(Xaa)n-SR; R: CH2CH2COOH,CH2C6H5, C6H4NHC(:NH)NH2), which proceeds efficiently in both a sequential manner and parallel to thesubsequent enzymatic reaction. The resulting substrate mimetics act as efficient acyl donor componentsand show the typical behavior of substrate mimicry enabling irreversible reactions with originally nonspecificacyl moieties. Neither a workup of the substrate mimetic intermediate nor changes of the reaction conditionsduring the whole synthesis process are required. Model peptide syntheses using trypsin, -chymotrypsin,and V8 protease as the biocatalysts proved the function of the approach and illustrated its synthetic valuefor protease-mediated reactions and the compatibility of the approach with state-of-the-art solid-phasepeptide ester synthesis methods.