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 Besir; Vladimir Benes; Juri Rappsilber; Martin Jechlinger; Jan O. Korbel; Imre Berger; Stefan Braese; Edward A. Lemke

doi:10.1038/nmeth.4032

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

Research output: Contribution to journal › Review article › peer-review

47 Scopus citations

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 language	English (US)
Pages (from-to)	997-1000
Number of pages	4
Journal	Nature Methods
Volume	13
Issue number	12
DOIs	https://doi.org/10.1038/nmeth.4032
State	Published - Dec 1 2016
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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10.1038/nmeth.4032

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Koehler, C., Sauter, P. F., Wawryszyn, M., Girona, G. E., Gupta, K., Landry, J. J. M., Fritz, M. H. Y., Radic, K., Hoffmann, J. E., Chen, Z. A., Zou, J., Tan, P. S., Galik, B., Junttila, S., Stolt-Bergner, P., Pruneri, G., Gyenesei, A., Schultz, C., Biskup, M. B., ... Lemke, E. A. (2016). Genetic code expansion for multiprotein complex engineering. Nature Methods, 13(12), 997-1000. https://doi.org/10.1038/nmeth.4032

Koehler, C, Sauter, PF, Wawryszyn, M, Girona, GE, Gupta, K, Landry, JJM, Fritz, MHY, Radic, K, Hoffmann, JE, Chen, ZA, Zou, J, Tan, PS, Galik, B, Junttila, S, Stolt-Bergner, P, Pruneri, G, Gyenesei, A, Schultz, C, Biskup, MB, Besir, H, Benes, V, Rappsilber, J, Jechlinger, M, Korbel, JO, Berger, I, Braese, S & Lemke, EA 2016, 'Genetic code expansion for multiprotein complex engineering', Nature Methods, vol. 13, no. 12, pp. 997-1000. https://doi.org/10.1038/nmeth.4032

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title = "Genetic code expansion for multiprotein complex engineering",

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 F{\"o}rster resonance energy transfer as well as site-specific crosslinking strategies.",

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

note = "Funding Information: We thank all members of our laboratories for helpful discussions. E.A.L., C.K., P.F.S., M.W., and S.B. acknowledge funding from the BW Stiftung. E.A.L. acknowledges additional support from the Emmy Noether program. E.A.L. and C.S. are grateful for funding by SPP1623 of the Deutsche Forschungsgemeinschaft. I.B. is funded by the European Commission Framework Programme 7 (FP7) ComplexINC project (contract no. 279039). P.S.-B. acknowledges funding from the Laura Bassi Centres of Expertise initiative for the Centre of Optimized Structural Studies, project 253275. M.W. thanks the KSOP for financial support. P.S.T. is supported by the EMBL Interdisciplinary Postdoc Programme (EIPOD) under Marie Curie Actions COFUND. The Wellcome Trust generously funded this work through a Senior Research Fellowship to J.R. (103139), a Centre core grant (092076), and an instrument grant (108504). We also thank the members of the EMBL Genomics Core Facility for sample processing and sequencing, as well as the EMBL FACS facility for technical support.",

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T1 - Genetic code expansion for multiprotein complex engineering

AU - Koehler, Christine

AU - Sauter, Paul F.

AU - Wawryszyn, Mirella

AU - Girona, Gemma Estrada

AU - Gupta, Kapil

AU - Landry, Jonathan J.M.

AU - Fritz, Markus Hsi Yang

AU - Radic, Ksenija

AU - Hoffmann, Jan Erik

AU - Chen, Zhuo A.

AU - Zou, Juan

AU - Tan, Piau Siong

AU - Galik, Bence

AU - Junttila, Sini

AU - Stolt-Bergner, Peggy

AU - Pruneri, Giancarlo

AU - Gyenesei, Attila

AU - Schultz, Carsten

AU - Biskup, Moritz Bosse

AU - Besir, Hueseyin

AU - Benes, Vladimir

AU - Rappsilber, Juri

AU - Jechlinger, Martin

AU - Korbel, Jan O.

AU - Berger, Imre

AU - Braese, Stefan

AU - Lemke, Edward A.

N1 - Funding Information: We thank all members of our laboratories for helpful discussions. E.A.L., C.K., P.F.S., M.W., and S.B. acknowledge funding from the BW Stiftung. E.A.L. acknowledges additional support from the Emmy Noether program. E.A.L. and C.S. are grateful for funding by SPP1623 of the Deutsche Forschungsgemeinschaft. I.B. is funded by the European Commission Framework Programme 7 (FP7) ComplexINC project (contract no. 279039). P.S.-B. acknowledges funding from the Laura Bassi Centres of Expertise initiative for the Centre of Optimized Structural Studies, project 253275. M.W. thanks the KSOP for financial support. P.S.T. is supported by the EMBL Interdisciplinary Postdoc Programme (EIPOD) under Marie Curie Actions COFUND. The Wellcome Trust generously funded this work through a Senior Research Fellowship to J.R. (103139), a Centre core grant (092076), and an instrument grant (108504). We also thank the members of the EMBL Genomics Core Facility for sample processing and sequencing, as well as the EMBL FACS facility for technical support.

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N2 - 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.

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ER -

Genetic code expansion for multiprotein complex engineering

Abstract

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