Inhibitory Mechanism of an Allosteric Antibody Targeting the Glucagon Receptor

Mukund, Susmith; Shang, Yonglei; Clarke, Holly J.; Madjidi, Azadeh; Corn, Jacob E.; Kates, Lance; Kolumam, Ganesh; Chiang, Vicky; Luis, Elizabeth; Murray, Jeremy; Zhang, Yingnan; Hötzel, Isidro; Koth, Christopher M.; Allan, Bernard B.

doi:10.1074/jbc.m113.496984

Inhibitory Mechanism of an Allosteric Antibody Targeting the Glucagon Receptor

Journal Article · Sun Nov 03 23:00:00 EST 2013 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.m113.496984· OSTI ID:1625083

Mukund, Susmith ^[1]; Shang, Yonglei ^[2]; Clarke, Holly J. ^[3]; Madjidi, Azadeh ^[3]; Corn, Jacob E. ^[4]; Kates, Lance ^[5]; Kolumam, Ganesh ^[5]; Chiang, Vicky ^[2]; Luis, Elizabeth ^[6]; Murray, Jeremy ^[7]; Zhang, Yingnan; Hötzel, Isidro ^[2]; Koth, Christopher M. ^[7]; Allan, Bernard B. ^[3]

Genentech, Inc., South San Francisco, CA (United States). Dept. of Structural Biology; DOE/OSTI
Genentech, Inc., South San Francisco, CA (United States). Dept. of Antibody Engineering
Genentech, Inc., South San Francisco, CA (United States). Dept. of Molecular Biology
Genentech, Inc., South San Francisco, CA (United States). Dept. of Early Discovery Biochemistry
Genentech, Inc., South San Francisco, CA (United States). Dept. of Biomedical Imaging
Genentech, Inc., South San Francisco, CA (United States). Dept. of Protein Chemistry
Genentech, Inc., South San Francisco, CA (United States). Dept. of Structural Biology

Elevated glucagon levels and increased hepatic glucagon receptor (GCGR) signaling contribute to hyperglycemia in type 2 diabetes. We have identified a monoclonal antibody that inhibits GCGR, a class B G-protein coupled receptor (GPCR), through a unique allosteric mechanism. Receptor inhibition is mediated by the binding of this antibody to two distinct sites that lie outside of the glucagon binding cleft. One site consists of a patch of residues that are surface-exposed on the face of the extracellular domain (ECD) opposite the ligand-binding cleft, whereas the second binding site consists of residues in the A helix of the ECD. A docking model suggests that the antibody does not occlude the ligand-binding cleft. We solved the crystal structure of GCGR ECD containing a naturally occurring G40S mutation and found a shift in the register of the A helix that prevents antibody binding. We also found that alterations in the A helix impact the normal function of GCGR. We present a model for the allosteric inhibition of GCGR by a monoclonal antibody that may form the basis for the development of allosteric modulators for the treatment of diabetes and other class B GPCR-related diseases.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1625083

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Journal Issue: 50 Vol. 288; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: English

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