Structure of avian AICAR transformylase with a multisubstrate adduct inhibitor β-DADF identifies the folate binding site

Dennis W. Wolan, Samantha E. Greasley, Mark J. Wall, Stephen Benkovic, Ian A. Wilson

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

The penultimate catalytic step of the purine de novo synthesis pathway is the conversion of aminoimidazole-4-carboxamide ribonucleotide (AICAR) to 5-formyl-AICAR that requires the cofactor N 10-formyl-tetrahydrofolate as the formyl donor. This reaction is catalyzed by the AICAR transformylase domain of the bifunctional enzyme AICAR transformylase/inosine monophosphate cyclohydrolase (ATIC). Identification of the location of the AICAR transformylase active site was previously elucidated from the crystal structure of the avian ATIC with bound substrate AICAR; however, due to the absence of any bound folate, the folate binding region of the active site could not be identified. Here, we have determined the homodimeric crystal structure of avian ATIC in complex with the ATIC-specific multisubstrate adduct inhibitor β-DADF to 2.5 Å resolution. β-DADF encompasses both the AICAR and folate moieties into a single covalently linked entity, thereby allowing for the characterization of the folate binding pocket of the AICAR transformylase active site. β-DADF is intimately bound at the dimer interface of the transformylase domains with the majority of AICAR moiety interactions occurring within one subunit, whereas the primary interactions to the folate occur with the opposing subunit. The crystal structure suggests that a buried Lys267 is transiently protonated during formyl transfer allowing for the stabilization of the oxyanion transition state and subsequent protonation of N10 on the tetrahydrofolate leaving group. Furthermore, the β-DADF-bound structure provides a more optimal three-dimensional scaffold to improve the design of specific antineoplastic agents.

Original languageEnglish (US)
Pages (from-to)10904-10914
Number of pages11
JournalBiochemistry
Volume42
Issue number37
DOIs
StatePublished - Sep 23 2003

All Science Journal Classification (ASJC) codes

  • Biochemistry

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