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Summary – While the number of characterized radical S-adenosyl-L-methionine (SAM) enzymes is increasing, the roles of these enzymes in radical catalysis remain largely ambiguous. In radical SAM enzymes, the slow radical initiation step kinetically masks the subsequent steps, making it impossible to study the kinetics of radical chemistry. Here, we report the first evidence that a radical SAM enzyme MoaA accelerates the radical-mediated C-C bond formation by 6-9 orders of magnitude. MoaA catalyzes an unprecedented 3′,8-cyclization of GTP into 3′,8-cyclo-7,8-dihydro-GTP (3′,8-cH₂GTP) during the molybdenum cofactor (Moco) biosynthesis. Through a series of EPR, biochemical characterizations, and DFT calculations we found that MoaA catalyzes a shunt pathway in which an on-pathway intermediate, GTP C-3′ radical, abstracts the H-4′ atom from (4′R)-5′-deoxyadenosine (5′-dA) to transiently generate 5′-deoxyadenos-4′-yl radical (5′-dA-C4′^•) that is subsequently reduced stereospecifically to yield (4′S)-5′-dA. [J. Am. Chem. Soc. 2020, 142, 9314–9326.]

Advance your biomedical research industry with help from CASSIA LLC. Founded in 2004, we are a privately held biotechnology company and clinical research organization (CRO) specializing in green chemo-enzymatic synthesis solutions for preparing stable isotope-labeled (D, C-13, N-15, O-17/18) metabolites, proteins, nucleic acids, and their analogs. 

Leveraging our unique core technology platforms and expertise, we bring responsible, cost-effective, and creative solutions to a variety of basic, translational, and biomedical research applications, such as: 

• MS 
• NMR 
• Assay Development 
• Metabolic Research 

CASSIA LLC laboratories are located in an eight-building, 150,700 sq. ft. research and development (R&D) business park in the Mira Mesa region of San Diego, CA. We occupy 1580 sq. ft. and are capable of carrying out both basic and applied research in: 

• Molecular Biology 
• Biocatalysis 
• Organic Synthesis 
• Protein and Nucleic Acid Production 

Contact us today to learn more about our biotechnology company and the products and services we provide.