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

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

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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 language	English
Pages (from-to)	997–1000
Number of pages	4
Journal	Nature Methods
Volume	13
Issue number	12
Early online date	17 Oct 2016
DOIs	https://doi.org/10.1038/nmeth.4032
Publication status	Published - Dec 2016

Keywords

Biophysical chemistry
Chemical tools
Genetic engineering
Protein design
Proteins

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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

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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.",

keywords = "Biophysical chemistry, Chemical tools, Genetic engineering, Protein design, Proteins",

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.}",

year = "2016",

month = dec,

doi = "10.1038/nmeth.4032",

language = "English",

volume = "13",

pages = "997–1000",

journal = "Nature Methods",

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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

TY - JOUR

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.

PY - 2016/12

Y1 - 2016/12

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.

AB - 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.

KW - Biophysical chemistry

KW - Chemical tools

KW - Genetic engineering

KW - Protein design

KW - Proteins

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U2 - 10.1038/nmeth.4032

DO - 10.1038/nmeth.4032

M3 - Article (Academic Journal)

C2 - 27749839

AN - SCOPUS:84991584172

SN - 1548-7091

VL - 13

SP - 997

EP - 1000

JO - Nature Methods

JF - Nature Methods

IS - 12

ER -

Genetic code expansion for multiprotein complex engineering

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Professor Imre Berger

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Genetic code expansion for multiprotein complex engineering

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