The discovery of a new antibody for BRIL-fused GPCR structure determination.

Scientific Reports
Hikaru MiyagiJun-Ichi Saito

Abstract

G-protein-coupled receptors (GPCRs)-the largest family of cell-surface membrane proteins-mediate the intracellular signal transduction of many external ligands. Thus, GPCRs have become important drug targets. X-ray crystal structures of GPCRs are very useful for structure-based drug design (SBDD). Herein, we produced a new antibody (SRP2070) targeting the thermostabilised apocytochrome b562 from Escherichia coli M7W/H102I/R106L (BRIL). We found that a fragment of this antibody (SRP2070Fab) facilitated the crystallisation of the BRIL-tagged, ligand bound GPCRs, 5HT1B and AT2R. Furthermore, the electron densities of the ligands were resolved, suggesting that SPR2070Fab is versatile and adaptable for GPCR SBDD. We anticipate that this new tool will significantly accelerate structure determination of other GPCRs and the design of small molecular drugs targeting them.

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Citations

Nov 15, 2020·Biophysics Reviews·Parijat SarkarAmitabha Chattopadhyay
Apr 27, 2021·Frontiers in Molecular Biosciences·Amara JabeenShoba Ranganathan
May 6, 2021·Acta Crystallographica. Section D, Structural Biology·Risako Tamura-SakaguchiTerukazu Nogi
May 15, 2021·Trends in Pharmacological Sciences·Haitao ZhangVadim Cherezov
Aug 2, 2021·Chemistry and Physics of Lipids·Parijat Sarkar, Amitabha Chattopadhyay

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Methods Mentioned

BETA
X-ray
dot blot
column chromatography
size exclusion chromatography
glycosylation

Software Mentioned

PISA
PHASER
REFMAC5
AREAIMOL
MOLREP
CCP4
CCP4mg
KAMO
FORMULATRIX

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