Dissemin is shutting down on January 1st, 2025

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Nature Research, Nature Methods, 12(13), p. 997-1000, 2016

DOI: 10.1038/nmeth.4032

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Genetic code expansion for multiprotein complex engineering

Journal article published in 2016 by Christine Koehler, Paul F. Sauter, Mirella Wawryszyn, Gemma Estrada Girona, G. Estrada Girona, K. Gupta J. J. M. Landry, Jonathan J. M. Landry ORCID, M. Hsi Yang Fritz, Markus Hsi-Yang Fritz, Ksenija Radic, Jan-Erik Hoffmann ORCID, Zhuo A. Chen, Juan Zou, P. Siong Tanv, Piau Siong Tan and other authors.
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

We present a baculovirus-based protein engineering method that enables site-specific introduction of unique functionalities in a eukaryotic protein complex recombinantly produced in insect cells. We demonstrate the versatility of this efficient and robust protein production platform, ‘MultiBacTATAG’, (i) for the fluorescent labeling of target proteins and biologics using click chemistries, (ii) for glycoengineering of antibodies, and (iii) for structure–function studies of novel eukaryotic complexes using single-molecule Förster resonance energy transfer as well as site-specific crosslinking strategies.