論文

査読有り
2017年10月

Affinity-Guided Oxime Chemistry for Selective Protein Acylation in Live Tissue Systems

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
  • Tomonori Tamura
  • ,
  • Zhining Song
  • ,
  • Kazuma Amaike
  • ,
  • Shin Lee
  • ,
  • Sifei Yin
  • ,
  • Shigeki Kiyonaka
  • ,
  • Itaru Hamachi

139
40
開始ページ
14181
終了ページ
14191
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1021/jacs.7b07339
出版者・発行元
AMER CHEMICAL SOC

Catalyst-mediated protein modification is a powerful approach for the imaging and engineering of natural proteins. We have previously developed affinity-guided 4-dimethylaminopyridine (AGD) chemistry as an efficient protein modification method using a catalytic acyl transfer reaction. However, because of the high electrophilicity of the thioester acyl donor molecule, AGD chemistry suffers from nonspecific reactions to proteins other than the target protein in crude biological environments, such as cell lysates, live cells, and tissue samples. To overcome this shortcoming, we here report a new acyl donor/organocatalyst system that allows more specific and efficient protein modification. In this method, a highly nucleophilic pyridinium oxime (PyOx) catalyst is conjugated to a ligand specific to the target protein. The ligand-tethered PyOx selectively binds to the target protein and facilitates the acyl transfer reaction of a mild electrophilic N-acyl-N-alkylsulfonamide acyl donor on the protein surface. We demonstrated that the new catalytic system, called AGOX (affinity-guided oxime) chemistry, can modify target proteins, both in test tubes and cell lysates, more selectively and efficiently than AGD chemistry. Low-background fluorescence labeling of the endogenous cell-membrane proteins, carbonic anhydrase XII and the folate receptor, in live cells allowed for the precise quantification of diffusion coefficients in the protein's native environment. Furthermore, the excellent biocompatibility and bioorthogonality of AGOX chemistry were demonstrated by the selective labeling of an endogenous neurotransmitter receptor in mouse brain slices, which are highly complicated tissue samples.

Web of Science ® 被引用回数 : 25

リンク情報
DOI
https://doi.org/10.1021/jacs.7b07339
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000413057100038&DestApp=WOS_CPL
ID情報
  • DOI : 10.1021/jacs.7b07339
  • ISSN : 0002-7863
  • Web of Science ID : WOS:000413057100038

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