Btk SH2-kinase interface is critical for allosteric kinase activation and its targeting inhibits B-cell neoplasms

Duarte D, Lamontanara A, La Sala G, Jeong S, Sohn Y, Panjkovich A, Georgeon S, Kükenshöner T, Marcaida M, Pojer F, De Vivo M, Svergun D, Kim H, Dal Peraro M, Hantschel O, (2019) DOI

SASDF53 – Bruton's Tyrosine Kinase - kinase domain

Bruton's tyrosine kinase, kinase domain
MWexperimental 29 kDa
MWexpected 32 kDa
VPorod 52 nm3
log I(s) 2.49×101 2.49×100 2.49×10-1 2.49×10-2
Bruton's tyrosine kinase, kinase domain small angle scattering data  s, nm-1
ln I(s)
Bruton's tyrosine kinase, kinase domain Guinier plot ln 2.50×101 Rg: 2.1 nm 0 (2.1 nm)-2 s2
(sRg)2I(s)/I(0)
Bruton's tyrosine kinase, kinase domain Kratky plot 1.104 0 3 sRg
p(r)
Bruton's tyrosine kinase, kinase domain pair distance distribution function Rg: 2.1 nm 0 Dmax: 6.8 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Bruton's tyrosine kinase, kinase domain DAMMIN model

log I(s)
 s, nm-1
Bruton's tyrosine kinase, kinase domain PDB model

Synchrotron SAXS data for kinase domain (amino acids 391-659) wild-type of human Bruton’s tyrosine kinase (Btk) in 20 mM Tris-HCl, 150 mM NaCl, 5% Glycerol, 1 mM TCEP, pH 7.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Dectris Pilatus 1M detector at a sample-detector distance of 2.849 m and at a wavelength of λ = 0.099 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.91 and 4.15 mg/ml were measured at 10°C. 10 successive 0.5 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve. The final scattering curve was analysed using PRIMUS software (ATSAS). The model depicts the DAMMIN ab initio reconstruction superimposed with the crystal structure 1K2P (PDB).

Bruton's tyrosine kinase, kinase domain (Btk-KD)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   31.8 kDa
 
UniProt   Q06187 (391-659)
Sequence   FASTA
 
PDB code   1K2P