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Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo

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Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo. / Cigler, Marko; Müller, Thorsten G; Horn-Ghetko, Daniel; von Wrisberg, Marie-Kristin; Fottner, Maximilian; Goody, Roger S; Itzen, Aymelt; Lang, Kathrin.

In: ANGEW CHEM INT EDIT, Vol. 56, No. 49, 04.12.2017, p. 15737-15741.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Cigler, M, Müller, TG, Horn-Ghetko, D, von Wrisberg, M-K, Fottner, M, Goody, RS, Itzen, A & Lang, K 2017, 'Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo', ANGEW CHEM INT EDIT, vol. 56, no. 49, pp. 15737-15741. https://doi.org/10.1002/anie.201706927

APA

Cigler, M., Müller, T. G., Horn-Ghetko, D., von Wrisberg, M-K., Fottner, M., Goody, R. S., Itzen, A., & Lang, K. (2017). Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo. ANGEW CHEM INT EDIT, 56(49), 15737-15741. https://doi.org/10.1002/anie.201706927

Vancouver

Cigler M, Müller TG, Horn-Ghetko D, von Wrisberg M-K, Fottner M, Goody RS et al. Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo. ANGEW CHEM INT EDIT. 2017 Dec 4;56(49):15737-15741. https://doi.org/10.1002/anie.201706927

Bibtex

@article{e29ae8e1202140cf91fc6102d3168a2f,
title = "Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo",
abstract = "The characterization of low-affinity protein complexes is challenging due to their dynamic nature. Here, we present a method to stabilize transient protein complexes in vivo by generating a covalent and conformationally flexible bridge between the interaction partners. A highly active pyrrolysyl tRNA synthetase mutant directs the incorporation of unnatural amino acids bearing bromoalkyl moieties (BrCnK) into proteins. We demonstrate for the first time that low-affinity protein complexes between BrCnK-containing proteins and their binding partners can be stabilized in vivo in bacterial and mammalian cells. Using this approach, we determined the crystal structure of a transient GDP-bound complex between a small G-protein and its nucleotide exchange factor. We envision that this approach will prove valuable as a general tool for validating and characterizing protein-protein interactions in vitro and in vivo.",
keywords = "Amino Acids, Amino Acyl-tRNA Synthetases, GTP-Binding Protein Regulators, GTP-Binding Proteins, Green Fluorescent Proteins, HEK293 Cells, Humans, Models, Molecular, Mutation, Protein Binding, Protein Stability, Journal Article, Research Support, Non-U.S. Gov't",
author = "Marko Cigler and M{\"u}ller, {Thorsten G} and Daniel Horn-Ghetko and {von Wrisberg}, Marie-Kristin and Maximilian Fottner and Goody, {Roger S} and Aymelt Itzen and Kathrin Lang",
note = "{\textcopyright} 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.",
year = "2017",
month = dec,
day = "4",
doi = "10.1002/anie.201706927",
language = "English",
volume = "56",
pages = "15737--15741",
journal = "ANGEW CHEM INT EDIT",
issn = "1433-7851",
publisher = "John Wiley and Sons Ltd",
number = "49",

}

RIS

TY - JOUR

T1 - Proximity-Triggered Covalent Stabilization of Low-Affinity Protein Complexes In Vitro and In Vivo

AU - Cigler, Marko

AU - Müller, Thorsten G

AU - Horn-Ghetko, Daniel

AU - von Wrisberg, Marie-Kristin

AU - Fottner, Maximilian

AU - Goody, Roger S

AU - Itzen, Aymelt

AU - Lang, Kathrin

N1 - © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

PY - 2017/12/4

Y1 - 2017/12/4

N2 - The characterization of low-affinity protein complexes is challenging due to their dynamic nature. Here, we present a method to stabilize transient protein complexes in vivo by generating a covalent and conformationally flexible bridge between the interaction partners. A highly active pyrrolysyl tRNA synthetase mutant directs the incorporation of unnatural amino acids bearing bromoalkyl moieties (BrCnK) into proteins. We demonstrate for the first time that low-affinity protein complexes between BrCnK-containing proteins and their binding partners can be stabilized in vivo in bacterial and mammalian cells. Using this approach, we determined the crystal structure of a transient GDP-bound complex between a small G-protein and its nucleotide exchange factor. We envision that this approach will prove valuable as a general tool for validating and characterizing protein-protein interactions in vitro and in vivo.

AB - The characterization of low-affinity protein complexes is challenging due to their dynamic nature. Here, we present a method to stabilize transient protein complexes in vivo by generating a covalent and conformationally flexible bridge between the interaction partners. A highly active pyrrolysyl tRNA synthetase mutant directs the incorporation of unnatural amino acids bearing bromoalkyl moieties (BrCnK) into proteins. We demonstrate for the first time that low-affinity protein complexes between BrCnK-containing proteins and their binding partners can be stabilized in vivo in bacterial and mammalian cells. Using this approach, we determined the crystal structure of a transient GDP-bound complex between a small G-protein and its nucleotide exchange factor. We envision that this approach will prove valuable as a general tool for validating and characterizing protein-protein interactions in vitro and in vivo.

KW - Amino Acids

KW - Amino Acyl-tRNA Synthetases

KW - GTP-Binding Protein Regulators

KW - GTP-Binding Proteins

KW - Green Fluorescent Proteins

KW - HEK293 Cells

KW - Humans

KW - Models, Molecular

KW - Mutation

KW - Protein Binding

KW - Protein Stability

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1002/anie.201706927

DO - 10.1002/anie.201706927

M3 - SCORING: Journal article

C2 - 28960788

VL - 56

SP - 15737

EP - 15741

JO - ANGEW CHEM INT EDIT

JF - ANGEW CHEM INT EDIT

SN - 1433-7851

IS - 49

ER -