Conformation of full-length Bruton tyrosine kinase (Btk) from synchrotron X-ray solution scattering.

Márquez JA, Smith CI, Petoukhov MV, Lo Surdo P, Mattsson PT, Knekt M, Westlund A, Scheffzek K, Saraste M, Svergun DI, EMBO J 22(18):4616-24 (2003) Europe PMC

SASDC52 – Bruton tyrosine kinase

Tyrosine-protein kinase BTK (R28C mutant)
MWI(0) 76 kDa
MWexpected 76 kDa
VPorod 130 nm3
log I(s) 1.83×102 1.83×101 1.83×100 1.83×10-1
Tyrosine-protein kinase BTK (R28C mutant) small angle scattering data  s, nm-1
ln I(s)
Tyrosine-protein kinase BTK (R28C mutant) Guinier plot ln 1.83×102 Rg: 5 nm 0 (5 nm)-2 s2
(sRg)2I(s)/I(0)
Tyrosine-protein kinase BTK (R28C mutant) Kratky plot 1.104 0 3 sRg
p(r)
Tyrosine-protein kinase BTK (R28C mutant) pair distance distribution function Rg: 5.6 nm 0 Dmax: 20 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Tyrosine-protein kinase BTK (R28C mutant) GASBOR model

log I(s)
 s, nm-1
Tyrosine-protein kinase BTK (R28C mutant) BUNCH model

Synchrotron SAXS data from solutions of Brutons's tyrosine kinase (Btk) in 20 mM HEPES, 200 mM NaCl, 2 mM DTT, 1 mM MgCl2, pH 7.5, were collected on the X33 beam line at the DORIS III storage ring (Hamburg, Germany) using a 1D Gas detector detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.15 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 10°C. 15 successive 60 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 and the different curves were scaled for protein concentration. 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 Guinier range is very short subject to experimental conditions

Tyrosine-protein kinase BTK (R28C mutant) (BTK)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   76.2 kDa
 
UniProt   Q06187
Sequence   FASTA