Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody

Andrii Ishchenko; Daniel Wacker; Mili Kapoor; Ai Zhang; Gye Won Han; Shibom Basu; Nilkanth Patel; Marc Messerschmidt; Uwe Weierstall; Wei Liu; Vsevolod Katritch; Bryan L. Roth; Raymond C. Stevens; Vadim Cherezov

doi:10.1073/pnas.1700891114

Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody

Andrii Ishchenko
, Daniel Wacker
, Mili Kapoor
, Ai Zhang
, Gye Won Han
, Shibom Basu
, Nilkanth Patel
, Marc Messerschmidt
, Uwe Weierstall
, Wei Liu
, Vsevolod Katritch
, Bryan L. Roth
, Raymond C. Stevens
, Vadim Cherezov

Research output: Contribution to journal › Article › peer-review

58 Scopus citations

Abstract

Monoclonal antibodies provide an attractive alternative to small-molecule therapies for a wide range of diseases. Given the importance of G protein-coupled receptors (GPCRs) as pharmaceutical targets, there has been an immense interest in developing therapeutic monoclonal antibodies that act on GPCRs. Here we present the 3.0-Å resolution structure of a complex between the human 5-hydroxy-tryptamine 2B (5-HT_2B) receptor and an antibody Fab fragment bound to the extracellular side of the receptor, determined by serial femtosecond crystallography with an X-ray free-electron laser. The antibody binds to a 3D epitope of the receptor that includes all three extracellular loops. The 5-HT_2B receptor is captured in a well-defined active-like state, most likely stabilized by the crystal lattice. The structure of the complex sheds light on the mechanism of selectivity in extracellular recognition of GPCRs by monoclonal antibodies.

Original language	English
Pages (from-to)	8223-8228
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	114
Issue number	31
DOIs	https://doi.org/10.1073/pnas.1700891114
State	Published - 1 Aug 2017
Externally published	Yes

Keywords

Active state
Antibody recognition
GPCR
Serial femtosecond crystallography
X-ray free-electron laser

Access to Document

10.1073/pnas.1700891114

Cite this

Ishchenko, A., Wacker, D., Kapoor, M., Zhang, A., Han, G. W., Basu, S., Patel, N., Messerschmidt, M., Weierstall, U., Liu, W., Katritch, V., Roth, B. L., Stevens, R. C., & Cherezov, V. (2017). Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody. Proceedings of the National Academy of Sciences of the United States of America, 114(31), 8223-8228. https://doi.org/10.1073/pnas.1700891114

@article{4773806339584f4a95a252d55f548fb9,

title = "Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody",

abstract = "Monoclonal antibodies provide an attractive alternative to small-molecule therapies for a wide range of diseases. Given the importance of G protein-coupled receptors (GPCRs) as pharmaceutical targets, there has been an immense interest in developing therapeutic monoclonal antibodies that act on GPCRs. Here we present the 3.0-{\AA} resolution structure of a complex between the human 5-hydroxy-tryptamine 2B (5-HT2B) receptor and an antibody Fab fragment bound to the extracellular side of the receptor, determined by serial femtosecond crystallography with an X-ray free-electron laser. The antibody binds to a 3D epitope of the receptor that includes all three extracellular loops. The 5-HT2B receptor is captured in a well-defined active-like state, most likely stabilized by the crystal lattice. The structure of the complex sheds light on the mechanism of selectivity in extracellular recognition of GPCRs by monoclonal antibodies.",

keywords = "Active state, Antibody recognition, GPCR, Serial femtosecond crystallography, X-ray free-electron laser",

author = "Andrii Ishchenko and Daniel Wacker and Mili Kapoor and Ai Zhang and Han, \{Gye Won\} and Shibom Basu and Nilkanth Patel and Marc Messerschmidt and Uwe Weierstall and Wei Liu and Vsevolod Katritch and Roth, \{Bryan L.\} and Stevens, \{Raymond C.\} and Vadim Cherezov",

note = "Funding Information: We thank S. Boutet for help with X-ray free-electron laser (XFEL) data collection, T. A. White for advice with XFEL data processing, M. Audet for advice with ELISA assays, and A. Walker for help with manuscript preparation. This work was supported by NIH Grants R01 GM108635 (to V.C.), R21 DA042298 (to W.L.), R01MH112205 (to B.L.R.), and U19MH82441 (to B.L.R.); NSF STC Award 1231306 (to U.W., W.L., M.M., and V.C.); the Center for Applied Structural Discovery at the Biodesign Institute at Arizona State University (U.W.); a National Institute of Mental Health Psychoactive Drug Screening Program contract (to B.L.R.); and the Michael Hooker Chair for Protein Therapeutics and Translational Proteomics (B.L.R.). Use of the Linac Coherent Light Source, SLAC National Accelerator Laboratory, was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract DE-AC02-76SF00515.",

year = "2017",

month = aug,

day = "1",

doi = "10.1073/pnas.1700891114",

language = "English",

volume = "114",

pages = "8223--8228",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "31",

}

Ishchenko, A, Wacker, D, Kapoor, M, Zhang, A, Han, GW, Basu, S, Patel, N, Messerschmidt, M, Weierstall, U, Liu, W, Katritch, V, Roth, BL, Stevens, RC & Cherezov, V 2017, 'Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody', Proceedings of the National Academy of Sciences of the United States of America, vol. 114, no. 31, pp. 8223-8228. https://doi.org/10.1073/pnas.1700891114

TY - JOUR

T1 - Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody

AU - Ishchenko, Andrii

AU - Wacker, Daniel

AU - Kapoor, Mili

AU - Zhang, Ai

AU - Han, Gye Won

AU - Basu, Shibom

AU - Patel, Nilkanth

AU - Messerschmidt, Marc

AU - Weierstall, Uwe

AU - Liu, Wei

AU - Katritch, Vsevolod

AU - Roth, Bryan L.

AU - Stevens, Raymond C.

AU - Cherezov, Vadim

N1 - Funding Information: We thank S. Boutet for help with X-ray free-electron laser (XFEL) data collection, T. A. White for advice with XFEL data processing, M. Audet for advice with ELISA assays, and A. Walker for help with manuscript preparation. This work was supported by NIH Grants R01 GM108635 (to V.C.), R21 DA042298 (to W.L.), R01MH112205 (to B.L.R.), and U19MH82441 (to B.L.R.); NSF STC Award 1231306 (to U.W., W.L., M.M., and V.C.); the Center for Applied Structural Discovery at the Biodesign Institute at Arizona State University (U.W.); a National Institute of Mental Health Psychoactive Drug Screening Program contract (to B.L.R.); and the Michael Hooker Chair for Protein Therapeutics and Translational Proteomics (B.L.R.). Use of the Linac Coherent Light Source, SLAC National Accelerator Laboratory, was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract DE-AC02-76SF00515.

PY - 2017/8/1

Y1 - 2017/8/1

N2 - Monoclonal antibodies provide an attractive alternative to small-molecule therapies for a wide range of diseases. Given the importance of G protein-coupled receptors (GPCRs) as pharmaceutical targets, there has been an immense interest in developing therapeutic monoclonal antibodies that act on GPCRs. Here we present the 3.0-Å resolution structure of a complex between the human 5-hydroxy-tryptamine 2B (5-HT2B) receptor and an antibody Fab fragment bound to the extracellular side of the receptor, determined by serial femtosecond crystallography with an X-ray free-electron laser. The antibody binds to a 3D epitope of the receptor that includes all three extracellular loops. The 5-HT2B receptor is captured in a well-defined active-like state, most likely stabilized by the crystal lattice. The structure of the complex sheds light on the mechanism of selectivity in extracellular recognition of GPCRs by monoclonal antibodies.

AB - Monoclonal antibodies provide an attractive alternative to small-molecule therapies for a wide range of diseases. Given the importance of G protein-coupled receptors (GPCRs) as pharmaceutical targets, there has been an immense interest in developing therapeutic monoclonal antibodies that act on GPCRs. Here we present the 3.0-Å resolution structure of a complex between the human 5-hydroxy-tryptamine 2B (5-HT2B) receptor and an antibody Fab fragment bound to the extracellular side of the receptor, determined by serial femtosecond crystallography with an X-ray free-electron laser. The antibody binds to a 3D epitope of the receptor that includes all three extracellular loops. The 5-HT2B receptor is captured in a well-defined active-like state, most likely stabilized by the crystal lattice. The structure of the complex sheds light on the mechanism of selectivity in extracellular recognition of GPCRs by monoclonal antibodies.

KW - Active state

KW - Antibody recognition

KW - GPCR

KW - Serial femtosecond crystallography

KW - X-ray free-electron laser

UR - https://www.scopus.com/pages/publications/85026770347

U2 - 10.1073/pnas.1700891114

DO - 10.1073/pnas.1700891114

M3 - Article

C2 - 28716900

AN - SCOPUS:85026770347

SN - 0027-8424

VL - 114

SP - 8223

EP - 8228

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 31

ER -

Structural insights into the extracellular recognition of the human serotonin 2B receptor by an antibody

Abstract

Keywords

Access to Document

Fingerprint

Cite this