Crystal Structure of G Protein-coupled Receptor Kinase 5 in Complex with a Rationally Designed Inhibitor

Abstract

Background: G protein-coupled receptor kinase 5 (GRK5), a cardiovascular disease target, has not been structurally characterized. Results: The 2.4 Å crystal structure of a GRK5⅐inhibitor complex was determined. Conclusion: Inhibitor confirms the rational design strategy, and GRK5 C-terminal region adopts a unique membrane-bound conformation. Significance: These results provide new insights into the design of selective inhibitors and how GRK4 subfamily members interact with membranes.

Small-Molecule G Protein–Coupled Receptor Kinase Inhibitors Attenuate G Protein–Coupled Receptor Kinase 2–Mediated Desensitization of Vasoconstrictor-Induced Arterial Contractions

“…These compounds, are 2-pyridylmethyl amide derivatives of GSK180736A (Waldschmidt et al, 2016), with CCG215022 being regarded as a 'pan' GRK inhibitor, with a high degree of selectivity for GRK over PKA (Homan et al, 2015), whereas CCG224063 shows high selectivity for GRK2/3 with >100 fold selectivity over GRK5, ROCK1, and PKA. In addition, we examined the ability of the previously characterised GRK2/3 inhibitor, Takeda compound 101, (3-[[[4-methyl-5-(4-pyridyl (Okawa et al, 2017;Thal et al, 2011) to inhibit UTP-mediated desensitization of arterial contractions.…”

Occludin S471 Phosphorylation Contributes to Epithelial Monolayer Maturation

“…GRK identified in this screen was inhibited in MDCK cells by using the kinasespecific inhibitors CCG215022 (GRK inhibitor 22 [GRK Inhib 22]) (32), 4-amino-5-(bromomethyl)-2-methylpyrimidine hydrobromide (ABMH; Santa Cruz), and paroxetine (Toronto Research Chemicals, Toronto, ON, Canada). Kinase assays with all three inhibitors were performed exactly as previously reported (32).…”

“…A list of potential kinases was compiled by entering the sequence around S471 (KELDDYREESEEYMAAADE) into three online kinase prediction programs: GPS (29), KinasePhos (30), and NetPhos (31). Kinases predicted to phosphorylate S471 by at least two of the three programs were included in an in vitro 32 P kinase assay (Millipore, Dundee, Scotland, United Kingdom) of the same peptide with two added lysines at the amino terminus to promote peptide capture on nitrocellulose filters (KKKELDDYREESEEYMAAADE; NeoBioSci, Cambridge, MA). GRK identified in this screen was inhibited in MDCK cells by using the kinasespecific inhibitors CCG215022 (GRK inhibitor 22 [GRK Inhib 22]) (32), 4-amino-5-(bromomethyl)-2-methylpyrimidine hydrobromide (ABMH; Santa Cruz), and paroxetine (Toronto Research Chemicals, Toronto, ON, Canada).…”

Structure and Function of the Hypertension Variant A486V of G Protein-coupled Receptor Kinase 4

“…Three recent structures of GRK5 bound to AMPPNP, sangivamycin, and an inhibitor (CCG215022) are also in press (30,31). The lack of a GRK4 structure hinders understanding of GRK4 function in particular and of the GRK family in general.…”

“…E, alignment of residues in the RH domain (␣0-␣1 helix, ␣9 helix, and C-terminal end of ␣11 helix) that form a hydrophobic domain-swap interface in GRK6 and GRK1 crystal structures (26,27,41) or a C-terminal/RH domain packing interface within monomeric GRK5 structures (30,31). Conserved residues in the domain-swap interface of GRK6-AMPPNP (26) are highlighted in yellow, and those also interacting in the GRK5 monomer structures (30,31) are in green. The LXXDL motif (residues 534 -538 of human GRK5) is conserved in GRK4␣ as described previously (30) and is underlined.…”