Structure‐Based Demystification of Radical Catalysis by a Coenzyme B<sub>12</sub> Dependent Enzyme—Crystallographic Study of Glutamate Mutase with Cofactor Homologues
Karl Gruber, Vanessa C. Csitkovits, Andrzej Łyskowski, Christoph Kratky, Bernhard Kräutler
Abstract
Abstract Catalysis by radical enzymes dependent on coenzyme B 12 (AdoCbl) relies on the reactive primary 5′‐deoxy‐5′adenosyl radical, which originates from reversible Co−C bond homolysis of AdoCbl. This bond homolysis is accelerated roughly 10 12 ‐fold upon binding the enzyme substrate. The structural basis for this activation is still strikingly enigmatic. As revealed here, a displaced firm adenosine binding cavity in substrate‐loaded glutamate mutase (GM) causes a structural misfit for intact AdoCbl that is relieved by the homolytic Co−C bond cleavage. Strategically interacting adjacent adenosine‐ and substrate‐binding protein cavities provide a tight caged radical reaction space, controlling the entire radical path. The GM active site is perfectly structured for promoting radical catalysis, including “negative catalysis”, a paradigm for AdoCbl‐dependent mutases.