unliganded and in complex with xyloglucan oligosaccharide. Enzyme displays an open active center groove grafted upon its beta-jelly roll fold, in which the side chaindecorations of xyloglucan may be accomodated. Binding of xylosyl and pendant galactosyl moieties is tolerated, but the enzyme is similarly competent in the binding of unbranched glucans
apo-CjGH74 comprising residues Pro35 to Ala765 and CjGH74 in complex with Glc4-based xyloglucooligosaccharides, as well as mutant enzymes D70A and D483A, vapour diffusion sitting drop method, from 0.1 M sodium acetate, pH 5.0, and 0.6 M sodium formate, 8% w/v PGA-LM with microseeding, combination of equal volumes of 5 mg/ml protein solution and mother liquor, 19°C, X-ray diffraction structure determination and analysis at 1.71-2.28 A resolution
purified enzyme free or in complex with substrate xyloglucan or inhibitor extracellular dermal glycoprotein, hanging drop vapor diffusion method, from 0.1 M sodium acetate, pH 5.5, and 1.5 M ammonium sulfate, or 0.1 M sodium acetate, pH 4.6, 25% PEG 3000, and 5 mg/ml digested xyloglucan, 20°C, X-ray diffraction structure determination and analysis at 0.95-2.70 A resolution
PoGH74cat as apoenzyme and as XLX complex, sitting drop vapour diffusion method, 0.0015 ml of 20 mg/ml protein with 10 mM xylose and 0.1 M glucose are mixed with 0.0015 ml of the reservoir solution 25% w/v PEG 8000, 100 mM Bis-Tris buffer, pH 6.9, 1 mM tris(2-carboxyethyl)phosphine, and 0.2 M sodium acetate, crystals of PoGH74cat in complex with XLX are grown by cocrystallization using 500 nl of 10 mg/ml protein plus 5 mM of a xyloglucan oligosaccharide mixture (XXXG, XLXG, XXLG, and XLLG) mixed with 500 nl of the reservoir solution 20% w/v PEG3350 and 0.2 M sodium potassium tartrate. Crystals of the PoGH74catD70A mutant in complex with XXLG and XGXXLG are obtained by cocrystallization of 500 nl of 10 mg/ml protein with a complex XyGO mixture mixed with 500 nl reservoir solution 25% w/v PEG3350, 0.2 M magnesium chloride, and 100 mM Tris buffer, pH 8.5, 22°C, X-ray diffraction structure determination and analysis at 1.50-2.10 A resolution, molecular replacement and modeling
unliganded and in complex with xyloglucan oligosaccharide. Enzyme displays an open active center groove grafted upon its (beta/alpha)8 fold, in which the side chaindecorations of xyloglucan may be accomodated. Kinetically productive interactions are made with both xylose and galactose substituents
loop mutant TmNXG1-DELTAYNIIG with an oligosaccharide product bound in the negative active-site subsites, X-ray diffraction structure determination and analysis at 2.0 A resolution, molecular replacement, molecular dynamics simulations and structure modelling of wild-type and mutant enzyme, detailed overview