Sugar binding to Kluyveromyces lactis glucokinase 1 (KlGlk1)

Renato H. Weiße.

SASDUH6 – Monomeric Kluyveromyces lactis glucokinase 1 triple point mutant (I356D, Y419D, H420D), 2.5 mg/mL, 10 mM fructose

Glucokinase-1 (H304Q)
MWexperimental 44 kDa
MWexpected 54 kDa
VPorod 76 nm3
log I(s) 3.03×10-2 3.03×10-3 3.03×10-4 3.03×10-5
Glucokinase-1 (H304Q) small angle scattering data  s, nm-1
ln I(s)
Glucokinase-1 (H304Q) Guinier plot ln 3.03×10-2 Rg: 2.5 nm 0 (2.5 nm)-2 s2
(sRg)2I(s)/I(0)
Glucokinase-1 (H304Q) Kratky plot 1.104 0 3 sRg
Dmax: 8.2 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Glucokinase-1 (H304Q) PYMOL model
Glucokinase-1 (H304Q) PYMOL model
Glucokinase-1 (H304Q) PYMOL model

Synchrotron SAXS data from solutions of Monomeric Kluyveromyces lactis glucokinase 1 triple point mutant (I356D, Y419D, H420D), 2.5 mg/mL, 10 mM fructose in 10 mM Tris-HCl, 1 mM DTT, 10 mM D-fructose, pH 7.4 were collected on the EMBL P12 beam line at the PETRA III storage ring (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.12398 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2.5 and 2.5 mg/ml were measured at 10°C. 30 successive 0.045 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.

Data set merged from three independent measurements of the same sample. CAUTION: Repulsive interparticle interference effects present in the sample result in inaccurate structural parameters (Rg, I(0), MW, Dmax) extracted from the data.

Glucokinase-1 (H304Q) (KlGlk1-H304Q)
Mol. type   Protein
Organism   Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37)
Olig. state   Monomer
Mon. MW   53.7 kDa
 
UniProt   Q6CUZ3 (1-481)
Sequence   FASTA