β(1–3)Glucan synthase ofNeurospora crassa: Reaction sequence based on kinetic evidence

Abstract

On the basis of the kinetic effects of substrate, activator, and inhibitors on β(1–3) glucan synthase activity ofNeurospora crassa, we propose the following reaction sequence for glucan synthesis. First, enzyme binds laminaribiose (activator), forming an enzyme-laminaribiose complex. Substrate (UDP-Glc) binding follows. UDP-Glc is hydrolyzed, releasing UDP, while the glucose residue remains associated with glucan synthase. The resulting enzyme-activator-glucose complex binds another UDP-Glc. It is likely that linear competitive inhibitors act at this step. Initial polymerization occurs, forming a disaccharide (which remains bound to glucan synthase) and UDP, which is released. The resulting enzyme-activator-disaccharide binds another UDP-Glc, and Glc is covalently added; further polymerization occurs by addition of Glc (from UDP-Glc) to the growing glucan chain, which remains associated with glucan synthase. Uncompetitive inhibitors are likely to affect enzyme activity at this step.