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, Nature Communications 11(1) (2020) DOI

SASDF73 – Bruton's Tyrosine Kinase - SH3-SH2-kinase domain

Bruton's tyrosine kinase - Src homology 3-2 kinase domain
MWexperimental 45 kDa
MWexpected 52 kDa
VPorod 72 nm3
log I(s) 4.42×101 4.42×100 4.42×10-1 4.42×10-2
Bruton's tyrosine kinase - Src homology 3-2 kinase domain small angle scattering data  s, nm-1
ln I(s)
Bruton's tyrosine kinase - Src homology 3-2 kinase domain Guinier plot ln 4.43×101 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Bruton's tyrosine kinase - Src homology 3-2 kinase domain Kratky plot 1.104 0 √3 sRg
p(r)
Bruton's tyrosine kinase - Src homology 3-2 kinase domain pair distance distribution function Rg: 2.6 nm 0 Dmax: 8.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Bruton's Tyrosine Kinase - SH3-SH2-kinase domain Rg histogram Rg, nm
log I(s)
 s, nm-1
Bruton's tyrosine kinase - Src homology 3-2 kinase domain DAMMIN model
log I(s)
 s, nm-1
Bruton's tyrosine kinase - Src homology 3-2 kinase domain PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data for auto-inhibited module SH3-SH2-kinase domain (amino acids 214-659) 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 0.78 and 4.32 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 raw curve measured from highest concentration was used for further analysis. The final scattering curve was analysed using PRIMUS software (ATSAS). The model depicts the DAMMIN ab initio reconstruction superimposed with the crystal structure 4XI2 (PDB). Ensemble optimisation method (EOM) was performed using three domains structure (1QLY, 2GE9 and 1K2P, PDB) to assess construct’s flexibility.

Bruton's tyrosine kinase - Src homology 3-2 kinase domain (hBTK - SH3-SH2-KD)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   52.2 kDa
 
UniProt   Q06187 (214-659)
Sequence   FASTA
 
PDB ID   4X12