Lc. Pedersen et al., Heparan/chondroitin sulfate biosynthesis - Structure and mechanism of human glucuronyltransferase I, J BIOL CHEM, 275(44), 2000, pp. 34580-34585
Human beta1,3-glucuronyltransferase I (GlcAT-I) is a central enzyme in the
initial steps of proteoglycan synthesis. GlcAT-I transfers a glucuronic aci
d moiety from the uridine diphosphate-glucuronic acid (UDP-GlcUA) to the co
mmon linkage region trisaccharide Gal beta1-3Gal beta1-4Xyl covalently boun
d to a Ser residue at the glycosaminylglycan attachment site of proteoglyca
ns, We have now determined the crystal structure of GlcAT-I at 2.3 Angstrom
in the presence of the donor substrate product UDP, the catalytic Mn2+ ion
, and the acceptor substrate analog Gal beta1-3Gal beta1-4Xyl, The enzyme i
s a alpha/beta protein with two subdomains that constitute the donor and ac
ceptor substrate binding site. The active site residues lie in a cleft exte
nding across both subdomains in which the trisaccharide molecule is oriente
d perpendicular to the UDP. Residues Glu(227), Asp(252), and Glu(281) dicta
te the binding orientation of the terminal Gal-2 moiety. Residue Glu(281) i
s in position to function as a catalytic base by deprotonating the incoming
3-hydroxyl group of the acceptor. The conserved DXD motif (Asp(191), Asp(1
95), Asp(196)) has direct interaction with the ribose of the UDP molecule a
s well as with the Mn2+ ion. The key residues involved in substrate binding
and catalysis are conserved in the glucuronyltransferase family as well as
of her glycosyltransferases.