Aminofutalosine Synthase (MqnE): A New Catalytic Motif in Radical SAM Enzymology. Chapter uri icon

abstract

  • Aminofutalosine synthase (MqnE) is a radical SAM enzyme involved in the futalosine-dependent menaquinone biosynthetic pathway. Its ability to add the 5'-deoxyadenosyl radical to the substrate-rather than abstract a hydrogen atom-and to catalyze radical addition to a stable benzene ring gives it a unique place in the radical SAM superfamily and required the development of new strategies for trapping radical intermediates. This chapter describes the methodologies used for enzyme overexpression, purification, and in vitro reconstitution. We also describe the development of fast, radical triggered, carbon-halogen bond fragmentation reactions for the trapping of intermediates. We anticipate that these methods will be of general use in the study of other transient enzymatic radicals.

author list (cited authors)

  • Joshi, S., Fedoseyenko, D., Mahanta, N., & Begley, T. P.

citation count

  • 9

complete list of authors

  • Joshi, Sumedh||Fedoseyenko, Dmytro||Mahanta, Nilkamal||Begley, Tadhg P

Book Title

  • RADICAL SAM ENZYMES

publication date

  • January 2018