4YHJ

Structure and Function of the Hypertension Variant A486V of G Protein-coupled Receptor Kinase 4 (GRK4)


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.60 Å
  • R-Value Free: 0.248 
  • R-Value Work: 0.197 
  • R-Value Observed: 0.200 

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This is version 1.3 of the entry. See complete history


Literature

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

Allen, S.J.Parthasarathy, G.Darke, P.L.Diehl, R.E.Ford, R.E.Hall, D.L.Johnson, S.A.Reid, J.C.Rickert, K.W.Shipman, J.M.Soisson, S.M.Zuck, P.Munshi, S.K.Lumb, K.J.

(2015) J Biol Chem 290: 20360-20373

  • DOI: https://doi.org/10.1074/jbc.M115.648907
  • Primary Citation of Related Structures:  
    4YHJ

  • PubMed Abstract: 

    G-protein-coupled receptor (GPCR) kinases (GRKs) bind to and phosphorylate GPCRs, initiating the process of GPCR desensitization and internalization. GRK4 is implicated in the regulation of blood pressure, and three GRK4 polymorphisms (R65L, A142V, and A486V) are associated with hypertension. Here, we describe the 2.6 Å structure of human GRK4α A486V crystallized in the presence of 5'-adenylyl β,γ-imidodiphosphate. The structure of GRK4α is similar to other GRKs, although slight differences exist within the RGS homology (RH) bundle subdomain, substrate-binding site, and kinase C-tail. The RH bundle subdomain and kinase C-terminal lobe form a strikingly acidic surface, whereas the kinase N-terminal lobe and RH terminal subdomain surfaces are much more basic. In this respect, GRK4α is more similar to GRK2 than GRK6. A fully ordered kinase C-tail reveals interactions linking the C-tail with important determinants of kinase activity, including the αB helix, αD helix, and the P-loop. Autophosphorylation of wild-type GRK4α is required for full kinase activity, as indicated by a lag in phosphorylation of a peptide from the dopamine D1 receptor without ATP preincubation. In contrast, this lag is not observed in GRK4α A486V. Phosphopeptide mapping by mass spectrometry indicates an increased rate of autophosphorylation of a number of residues in GRK4α A486V relative to wild-type GRK4α, including Ser-485 in the kinase C-tail.


  • Organizational Affiliation

    From Screening and Protein Sciences, Merck Research Laboratories, North Wales, Pennsylvania 19454 and samantha_allen@merck.com.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
G protein-coupled receptor kinase 4
A, B
584Homo sapiensMutation(s): 2 
Gene Names: GRK4GPRK2LGPRK4
EC: 2.7.11.16
UniProt & NIH Common Fund Data Resources
Find proteins for P32298 (Homo sapiens)
Explore P32298 
Go to UniProtKB:  P32298
PHAROS:  P32298
GTEx:  ENSG00000125388 
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP32298
Sequence Annotations
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  • Reference Sequence
Small Molecules
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.60 Å
  • R-Value Free: 0.248 
  • R-Value Work: 0.197 
  • R-Value Observed: 0.200 
  • Space Group: P 31 2 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 104.838α = 90
b = 104.838β = 90
c = 221.838γ = 120
Software Package:
Software NamePurpose
HKL-2000data collection
SCALEPACKdata scaling
BUSTER-TNTrefinement
PDB_EXTRACTdata extraction
HKL-2000data reduction
PHASERphasing

Structure Validation

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Ligand Structure Quality Assessment 


Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2015-07-08
    Type: Initial release
  • Version 1.1: 2015-07-15
    Changes: Database references
  • Version 1.2: 2015-09-02
    Changes: Database references
  • Version 1.3: 2024-02-28
    Changes: Data collection, Database references, Derived calculations