Genetic code expansion for multiprotein complex engineering

Christine Koehler*, Paul F. Sauter, Mirella Wawryszyn, Gemma Estrada Girona, Kapil Gupta, Jonathan J M Landry, Markus Hsi Yang Fritz, Ksenija Radic, Jan Erik Hoffmann, Zhuo A. Chen, Juan Zou, Piau Siong Tan, Bence Galik, Sini Junttila, Peggy Stolt-Bergner, Giancarlo Pruneri, Attila Gyenesei, Carsten Schultz, Moritz Bosse Biskup, Hueseyin BesirVladimir Benes, Juri Rappsilber, Martin Jechlinger, Jan O. Korbel, Imre Berger, Stefan Braese, Edward A. Lemke

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

39 Citations (Scopus)
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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, 'MultiBacTAG', (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.
Original languageEnglish
Pages (from-to)997–1000
Number of pages4
JournalNature Methods
Volume13
Issue number12
Early online date17 Oct 2016
DOIs
Publication statusPublished - Dec 2016

Keywords

  • Biophysical chemistry
  • Chemical tools
  • Genetic engineering
  • Protein design
  • Proteins

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