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Efficient and wash-free labeling of membrane proteins using engineered Npu DnaE split-inteins.

ABSTRACT: 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.

SUBMITTER: Lee E 

PROVIDER: S-EPMC6194237 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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Efficient and wash-free labeling of membrane proteins using engineered Npu DnaE split-inteins.

Lee Euiyeon E   Min Kyoungmi K   Chang Young-Tae YT   Kwon Youngeun Y  

Protein science : a publication of the Protein Society 20180924 9

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. ...[more]

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