Conformational heterogeneity of the BTK PHTH domain drives multiple regulatory states.

Lin DY, Kueffer LE, Juneja P, Wales TE, Engen JR, Andreotti AH, Elife 12 (2024) Europe PMC

SASDRB9 – Bruton's Tyrosine Kinase - full-length

Tyrosine-protein kinase BTK
MWI(0) 89 kDa
MWexpected 77 kDa
VPorod 118 nm3
log I(s) 8.91×101 8.91×100 8.91×10-1 8.91×10-2
Tyrosine-protein kinase BTK small angle scattering data  s, nm-1
ln I(s)
Tyrosine-protein kinase BTK Guinier plot ln 8.92×101 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
Tyrosine-protein kinase BTK Kratky plot 1.104 0 √3 sRg
p(r)
Tyrosine-protein kinase BTK pair distance distribution function Rg: 4.5 nm 0 Dmax: 19.1 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Tyrosine-protein kinase BTK DAMMIF model
Synchrotron SAXS data from solutions of Bruton's tyrosine kinase in 20 mM Tris-HCl, 150 mM NaCl, 1 mM DTT, 2% v/v glycerol, pH 8 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS, Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 1 and 10 mg/ml were measured at 20.9°C. 50 successive 0.200 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 concentrations were extrapolated to infinite dilution and merged with the higher concentration data to yield the final composite scattering curve.

Tyrosine-protein kinase BTK (Btk)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   77.2 kDa
 
UniProt   P35991 (1-659)
Sequence   FASTA