title: Screening-based discovery and structural dissection of a novel family 18 chitinase inhibitor.
abstract: Family 18 chitinases play key roles in the life cycles of a variety of organisms ranging from bacteria to man. Very recently it has been shown that one of the mammalian chitinases is highly overexpressed in the asthmatic lung and contributes to the pathogenic process through recruitment of inflammatory cells. Although several potent natural product chitinase inhibitors have been identified, their chemotherapeutic potential or their use as cell biological tools is limited due to their size, complex chemistry, and limited availability. We describe a virtual screening-based approach to identification of a novel, purine-based, chitinase inhibitor. This inhibitor acts in the low micromolar (Ki=2.8+/-0.2 microM) range in a competitive mode. Dissection of the binding mode by x-ray crystallography reveals that the compound, which consists of two linked caffeine moieties, binds in the active site through extensive and not previously observed stacking interactions with conserved, solvent exposed tryptophans. Such exposed aromatics are also present in the structures of many other carbohydrate processing enzymes. The compound exhibits favorable chemical properties and is likely to be useful as a general scaffold for development of pan-family 18 chitinase inhibitors.
authors.affiliation: Division of Biological Chemistry and Molecular Microbiology, Wellcome Trust Biocentre, School of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland.
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Schüttelkopf
authors.forename: Ole A
authors.initials: OA
authors.lastname: Andersen
authors.forename: Francesco V
authors.initials: FV
authors.lastname: Rao
authors.forename: Matthew
authors.initials: M
authors.lastname: Allwood
authors.forename: Clare
authors.initials: C
authors.lastname: Lloyd
authors.forename: Ian M
authors.initials: IM
authors.lastname: Eggleston
authors.forename: Daan M F
authors.initials: DM
authors.lastname: van Aalten
mesh_terms: D000465:Algorithms; D001232:Aspergillus fumigatus / Q000201:enzymology; D001665:Binding Sites; D001671:Biochemistry / Q000379:methods*; D002110:Caffeine / Q000737:chemistry; D002688:Chitinases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D019295:Computational Biology / Q000379:methods; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D014364:Tryptophan / Q000737:chemistry
chemical_list: D004791:Enzyme Inhibitors; D002110:Caffeine; D014364:Tryptophan; D002688:Chitinases