title: The antiviral drug acyclovir is a slow-binding inhibitor of (D)-amino acid oxidase.
abstract: d-Amino acid oxidase (DAO) is a degradative enzyme that is stereospecific for d-amino acids, including d-serine and d-alanine, which are believed to be coagonists of the N-methyl-d-aspartate (NMDA) receptor. To identify a new class of DAO inhibitor(s) that can be used to elucidate the molecular details of the active site environment of DAO, manifold biologically active compounds of microbial origin and pre-existing drugs were screened for their ability to inhibit DAO activity, and several compounds were identified as candidates. One of these compounds, acyclovir (ACV), a well-known antiviral drug used for the treatment of herpesvirus infections, was characterized and evaluated as a novel DAO inhibitor in vitro. Analysis showed that ACV acts on DAO as a reversible slow-binding inhibitor, and interestingly, the time required to achieve equilibrium between DAO, ACV, and the DAO/ACV complex was highly dependent on temperature. The binding mechanism of ACV to DAO was investigated in detail by several approaches, including kinetic analysis, structural modeling of DAO complexed with ACV, and site-specific mutagenesis of an active site residue postulated to be involved in the binding of ACV. The results confirm that ACV is a novel, active site-directed inhibitor of DAO that can be a valuable tool for investigating the structure-function relationships of DAO, including the molecular details of the active site environment of DAO. In particular, it appears that ACV can serve as an active site probe to study the structural basis of temperature-induced conformational changes of DAO.
authors.affiliation: Laboratory of Biomolecular Science, Graduate School of Pharmaceutical Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
authors.forename: Masumi
authors.initials: M
authors.lastname: Katane
authors.forename: Satsuki
authors.initials: S
authors.lastname: Matsuda
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Saitoh
authors.forename: Masae
authors.initials: M
authors.lastname: Sekine
authors.forename: Takemitsu
authors.initials: T
authors.lastname: Furuchi
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Koyama
authors.forename: Izumi
authors.initials: I
authors.lastname: Nakagome
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tomoda
authors.forename: Shuichi
authors.initials: S
authors.lastname: Hirono
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Homma
mesh_terms: D000212:Acyclovir / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D000465:Algorithms; D000998:Antiviral Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001565:Benzoates / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D020134:Catalytic Domain / Q000235:genetics; D003605:D-Amino-Acid Oxidase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D013696:Temperature
chemical_list: D000998:Antiviral Agents; D001565:Benzoates; D003605:D-Amino-Acid Oxidase; D000212:Acyclovir