2IVU

Crystal structure of phosphorylated RET tyrosine kinase domain complexed with the inhibitor ZD6474


Experimental Data Snapshot

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.50 Å
  • R-Value Free: 0.248 
  • R-Value Work: 0.190 
  • R-Value Observed: 0.193 

wwPDB Validation   3D Report Full Report


This is version 2.3 of the entry. See complete history


Literature

Structure and chemical inhibition of the RET tyrosine kinase domain.

Knowles, P.P.Murray-Rust, J.Kjaer, S.Scott, R.P.Hanrahan, S.Santoro, M.Ibanez, C.F.McDonald, N.Q.

(2006) J Biol Chem 281: 33577-33587

  • DOI: https://doi.org/10.1074/jbc.M605604200
  • Primary Citation of Related Structures:  
    2IVS, 2IVT, 2IVU, 2IVV

  • PubMed Abstract: 

    The RET proto-oncogene encodes a receptor tyrosine kinase for the glial cell line-derived neurotrophic factor family of ligands. Loss-of-function mutations in RET are implicated in Hirschsprung disease, whereas activating mutations in RET are found in human cancers, including familial medullar thyroid carcinoma and multiple endocrine neoplasias 2A and 2B. We report here the biochemical characterization of the human RET tyrosine kinase domain and the structure determination of the non-phosphorylated and phosphorylated forms. Both structures adopt the same active kinase conformation competent to bind ATP and substrate and have a pre-organized activation loop conformation that is independent of phosphorylation status. In agreement with the structural data, enzyme kinetic data show that autophosphorylation produces only a modest increase in activity. Longer forms of RET containing the juxtamembrane domain and C-terminal tail exhibited similar kinetic behavior, implying that there is no cis-inhibitory mechanism within the RET intracellular domain. Our results suggest the existence of alternative inhibitory mechanisms, possibly in trans, for the autoregulation of RET kinase activity. We also present the structures of the RET tyrosine kinase domain bound to two inhibitors, the pyrazolopyrimidine PP1 and the clinically relevant 4-anilinoquinazoline ZD6474. These structures explain why certain multiple endocrine neoplasia 2-associated RET mutants found in patients are resistant to inhibition and form the basis for design of more effective inhibitors.


  • Organizational Affiliation

    Structural Biology Laboratory, London Research Institute, Cancer Research UK, London WC2A 3PX, UK.


Macromolecules
Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
PROTO-ONCOGENE TYROSINE-PROTEIN KINASE RECEPTOR RET PRECURSOR314Homo sapiensMutation(s): 0 
EC: 2.7.10.1
UniProt & NIH Common Fund Data Resources
Find proteins for P07949 (Homo sapiens)
Explore P07949 
Go to UniProtKB:  P07949
PHAROS:  P07949
GTEx:  ENSG00000165731 
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupP07949
Sequence Annotations
Expand
  • Reference Sequence
Small Molecules
Ligands 2 Unique
IDChains Name / Formula / InChI Key2D Diagram3D Interactions
ZD6
Query on ZD6

Download Ideal Coordinates CCD File 
D [auth A]Vandetanib
C22 H24 Br F N4 O2
UHTHHESEBZOYNR-UHFFFAOYSA-N
FMT
Query on FMT

Download Ideal Coordinates CCD File 
B [auth A],
C [auth A]
FORMIC ACID
C H2 O2
BDAGIHXWWSANSR-UHFFFAOYSA-N
Modified Residues  1 Unique
IDChains TypeFormula2D DiagramParent
PTR
Query on PTR
A
L-PEPTIDE LINKINGC9 H12 N O6 PTYR
Binding Affinity Annotations 
IDSourceBinding Affinity
ZD6 Binding MOAD:  2IVU Ki: 375 (nM) from 1 assay(s)
PDBBind:  2IVU Ki: 375 (nM) from 1 assay(s)
BindingDB:  2IVU Kd: min: 14, max: 1.00e+4 (nM) from 3 assay(s)
IC50: min: 4, max: 5000 (nM) from 12 assay(s)
Experimental Data & Validation

Experimental Data

  • Method: X-RAY DIFFRACTION
  • Resolution: 2.50 Å
  • R-Value Free: 0.248 
  • R-Value Work: 0.190 
  • R-Value Observed: 0.193 
  • Space Group: C 1 2 1
Unit Cell:
Length ( Å )Angle ( ˚ )
a = 71.409α = 90
b = 71.42β = 101.23
c = 78.834γ = 90
Software Package:
Software NamePurpose
REFMACrefinement
MOSFLMdata reduction
SCALAdata scaling
MOLREPphasing

Structure Validation

View Full Validation Report



Entry History 

Deposition Data

Revision History  (Full details and data files)

  • Version 1.0: 2006-08-10
    Type: Initial release
  • Version 1.1: 2011-05-08
    Changes: Version format compliance
  • Version 1.2: 2011-07-13
    Changes: Version format compliance
  • Version 1.3: 2017-06-28
    Changes: Data collection
  • Version 2.0: 2017-10-25
    Changes: Atomic model, Database references, Structure summary
  • Version 2.1: 2018-02-28
    Changes: Database references
  • Version 2.2: 2019-01-30
    Changes: Data collection, Experimental preparation
  • Version 2.3: 2019-05-08
    Changes: Data collection, Derived calculations, Experimental preparation