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

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 use...

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Bibliographic Details
Published in:Journal of the American Chemical Society 2017-05, Vol.139 (17), p.6146-6151
Main Authors: Zhang, Yingqian, Zhou, Xiaoping, Xie, Yonghui, Greenberg, Marc M, Xi, Zhen, Zhou, Chuanzheng
Format: Article
Language:English
Subjects:
Acetolactate Synthase - chemistry
Acetolactate Synthase - metabolism
Biocompatible Materials - chemistry
Biocompatible Materials - metabolism
Escherichia coli - enzymology
Isoenzymes - chemistry
Isoenzymes - metabolism
Models, Molecular
Molecular Structure
Pyrroles - chemical synthesis
Pyrroles - chemistry
Pyrroles - metabolism
Sulfhydryl Compounds - chemistry
Sulfhydryl Compounds - metabolism
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Summary: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.
ISSN:0002-7863
1520-5126
DOI:10.1021/jacs.7b00670