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Efficient and wash-free labeling of membrane proteins using engineered Npu DnaE split-inteins
- Lee, Euiyeon;
- Min, Kyoungmi;
- Chang, Young-Tae;
- Kwon, Youngeun
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5초록
An efficient and wash-free method to conjugate a fluorescent tag to a target membrane protein is developed, using engineered Npu DnaE split-inteins. This approach allowed fast labeling while avoiding the strenuous synthesis of a long polypeptide. Two different modes of labeling, namely specific binding and covalent conjugation, are observed. The covalent labeling was monitored within 5 min, without background staining.
키워드
split-intein; protein labeling; protein trans-splicing; protein engineering; CELL-SURFACE PROTEINS; NOSTOC-PUNCTIFORME; LIVE CELLS; IDENTIFICATION; ACTIVATION; BIOSENSOR; DESIGN; TAGS
- 제목
- Efficient and wash-free labeling of membrane proteins using engineered Npu DnaE split-inteins
- 저자
- Lee, Euiyeon; Min, Kyoungmi; Chang, Young-Tae; Kwon, Youngeun
- DOI
- 10.1002/pro.3455
- 발행일
- 2018-09
- 유형
- Article
- 저널명
- Protein Science
- 권
- 27
- 호
- 9
- 페이지
- 1568 ~ 1574