| Autor: |
Qu W, Catcott KC, Zhang K; School of Pharmaceutical Science and Technology, Tianjin University , Tianjin 300072, China., Liu S, Guo JJ, Ma J; School of Pharmaceutical Science, Wenzhou Medical University , Wenzhou 325035, China., Pablo M, Glick J, Xiu Y, Kenton N, Ma X, Duclos RI Jr, Zhou ZS |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of the American Chemical Society [J Am Chem Soc] 2016 Mar 09; Vol. 138 (9), pp. 2877-80. Date of Electronic Publication: 2016 Feb 29. |
| DOI: |
10.1021/jacs.5b05950 |
| Abstrakt: |
Identifying an enzyme's substrates is essential to understand its function, yet it remains challenging. A fundamental impediment is the transient interactions between an enzyme and its substrates. In contrast, tight binding is often observed for multisubstrate-adduct inhibitors due to synergistic interactions. Extending this venerable concept to enzyme-catalyzed in situ adduct formation, unknown substrates were affinity-captured by an S-adenosyl-methionine (AdoMet, SAM)-dependent methyltransferase (MTase). Specifically, the electrophilic methyl sulfonium (alkyl donor) in AdoMet is replaced with a vinyl sulfonium (Michael acceptor) in S-adenosyl-vinthionine (AdoVin). Via an addition reaction, AdoVin and the nucleophilic substrate form a covalent bisubstrate-adduct tightly complexed with thiopurine MTase (2.1.1.67). As such, an unknown substrate was readily identified from crude cell lysates. Moreover, this approach is applicable to other systems, even if the enzyme is unknown. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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