Synchrotron SAXS data from solutions of Heparin oligosaccharide DP12 in 50 mM Tris/HCl, 60 mM CaCl2, pH 7.4 were collected on the BM29 beam line at the ESRF storage ring (Grenoble, France) using a Pilatus3 2M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured at 20°C. 10 successive 0.500 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.
Serial dilutions from 5 mg/ml to 1.25 mg/ml were collected under flow with the sample changer. At 60 mM Ca2+ aggregation started to appear. These oligosaccharides have been prepared by high resolution gel filtration of partial heparin lyase (Heparinase I) digestion of high quality porcin heparin.
General formula*
∆HexA,2S - GlcNS,6S– (IdoUA,2S – GlcNS,6S)n
n = number of disaccharide units.
*Although the main disaccharide unit in these products is IdoUA,2S – GlcNS,6S, (approx 75%) saccharides in each size class show some variation in degree and pattern of sulphation.
Uronic acid (HexA) at the non-reducing end of the oligosaccharides has a C4-C5 double bond as a result of the endolytic action of bacterial heparin lyase.
s, nm-1
