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

SASDRD9 – Bruton's Tyrosine Kinase - SH3-SH2-kinase domain

Tyrosine-protein kinase BTK (SH3-SH2-kinase domains)
MWexperimental 46 kDa
MWexpected 52 kDa
VPorod 68 nm3
log I(s) 2.62×101 2.62×100 2.62×10-1 2.62×10-2
Tyrosine-protein kinase BTK (SH3-SH2-kinase domains) small angle scattering data  s, nm-1
ln I(s)
Tyrosine-protein kinase BTK (SH3-SH2-kinase domains) Guinier plot ln 2.63×101 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
Tyrosine-protein kinase BTK (SH3-SH2-kinase domains) Kratky plot 1.104 0 √3 sRg
p(r)
Tyrosine-protein kinase BTK (SH3-SH2-kinase domains) pair distance distribution function Rg: 2.5 nm 0 Dmax: 7.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Tyrosine-protein kinase BTK (SH3-SH2-kinase domains) DAMMIF model
Synchrotron SAXS data from solutions of the SH3-SH2-kinase domain from 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 5 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 (SH3-SH2-kinase domains) (Btk)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   52.5 kDa
 
UniProt   P35991 (214-659)
Sequence   FASTA