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

SASDF83 – Bruton's Tyrosine Kinase - Full-length

Bruton's tyrosine kinase - full length
MWexperimental 67 kDa
MWexpected 77 kDa
VPorod 114 nm3
log I(s) 6.02×101 6.02×100 6.02×10-1 6.02×10-2
Bruton's tyrosine kinase - full length small angle scattering data  s, nm-1
ln I(s)
Bruton's tyrosine kinase - full length Guinier plot ln 6.02×101 Rg: 4.0 nm 0 (4.0 nm)-2 s2
Bruton's tyrosine kinase - full length Kratky plot 1.104 0 3 sRg
Bruton's tyrosine kinase - full length pair distance distribution function Rg: 4.3 nm 0 Dmax: 15.6 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Bruton's Tyrosine Kinase - Full-length Rg histogram Rg, nm

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

log I(s)
 s, nm-1
Bruton's tyrosine kinase - full length SREFLEX model

Synchrotron SAXS data for full-length (amino acids 1-659) 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 0.42 and 5.48 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 atomic model SASDC52 (EMBL, Hamburg) subject to SREFLEX to improve its agreement with experimental data. Ensemble optimisation method (EOM) was performed using all four domains structure (2Z0P, 1QLY, 2GE9 and 1K2P, PDB) to assess construct’s flexibility.

Bruton's tyrosine kinase - full length (hBTK - FL)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   76.7 kDa
UniProt   Q06187 (1-659)
Sequence   FASTA