2. Academic Validation
  3. Thiol Specific and Tracelessly Removable Bioconjugation via Michael Addition to 5-Methylene Pyrrolones

Thiol Specific and Tracelessly Removable Bioconjugation via Michael Addition to 5-Methylene Pyrrolones

  • J Am Chem Soc. 2017 May 3;139(17):6146-6151. doi: 10.1021/jacs.7b00670.
Yingqian Zhang 1,  Xiaoping Zhou 1,  Yonghui Xie 1,  Marc M Greenberg 2,  Zhen Xi 1 3,  Chuanzheng Zhou 1 3
Affiliations

Affiliations

  • 1 State Key Laboratory of Elemento-Organic Chemistry and Department of Chemical Biology, College of Chemistry, Nankai University , Tianjin 300071, China.
  • 2 Department of Chemistry, Johns Hopkins University , 3400 North Charles Street, Baltimore, Maryland 21218, United States.
  • 3 Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300071, China.
PMID: 28407468 DOI: 10.1021/jacs.7b00670
Abstract

5-Methylene pyrrolones (5MPs) are highly thiol-specific and tracelessly removable bioconjugation tools. 5MPs are readily prepared from primary amines in one step. 5MPs exhibit significantly improved stability under physiologically relevant conditions and cysteine specificity compared to commonly used analogues, maleimides. Michael addition of thiol to 5MPs occurs rapidly, cleanly, and does not generate a stereocenter. The conjugates efficiently release thiols via retro-Michael reaction in alkaline buffer (pH 9.5) or via thiol exchange at pH 7.5. This unique property makes 5MPs valuable for the controlled release of conjugated cargo and temporary thiol protection. The utilization of 5MPs for protein immobilization and pull-down of active complexes is illustrated using E. coli. acetohydroxyacid synthase isozyme I.

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