Binding of 5-GTP to the C-terminal FeS Cluster of the Radical S-Adenosylmethionine Enzyme MoaA Provides Insights into its Mechanism

Journal Article · Sun Jan 01 04:00:00 EST 2006 · Proc Natl Acad Sci USA

DOI:https://doi.org/10.1073/pnas.0510711103· OSTI ID:914306

Haenzelmann, P; Schindelin, H

The first step in molybdenum cofactor biosynthesis, the conversion of 5'-GTP to precursor Z, an oxygen-sensitive tetrahydropyranopterin is catalyzed by the S-adenosylmethionine (SAM)-dependent enzyme MoaA and the accessory protein MoaC. This reaction involves the radical-initiated intramolecular rearrangement of the guanine C8 atom. MoaA harbors an N-terminal [4Fe-4S] cluster, which is involved in the reductive cleavage of SAM and generates a 5'-deoxyadenosyl radical (5'-dA), and a C-terminal [4Fe-4S] cluster presumably involved in substrate binding and/or activation. Biochemical studies identified residues involved in 5'-GTP binding and the determinants of nucleotide specificity. The crystal structure of MoaA in complex with 5'-GTP confirms the biochemical data and provides valuable insights into the subsequent radical reaction. MoaA binds 5'-GTP with high affinity and interacts through its C-terminal [4Fe-4S] cluster with the guanine N1 and N2 atoms, in a yet uncharacterized binding mode. The tightly anchored triphosphate moiety prevents the escape of radical intermediates. This structure also visualizes the L-Met and 5'-dA cleavage products of SAM. Rotation of the 5'-dA ribose and/or conformational changes of the guanosine are proposed to bring the 5'-deoxyadenosyl radical into close proximity of either the ribose C2' and C3' or the guanine C8 carbon atoms leading to hydrogen abstraction.

• Research Organization: Brookhaven National Laboratory (BNL) National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: AC02-98CH10886
• OSTI ID: 914306
• Report Number(s): BNL--78874-2007-JA
• Journal Information: Proc Natl Acad Sci USA, Journal Name: Proc Natl Acad Sci USA Vol. 103; ISSN PNASA6; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

36 MATERIALS SCIENCE
BIOSYNTHESIS
CONFORMATIONAL CHANGES
CRYSTAL STRUCTURE
ENZYMES
GUANINE
MOLYBDENUM
RADICALS
national synchrotron light source