title: Preparation of allosamidin and demethylallosamidin photoaffinity probes and analysis of allosamidin-binding proteins in asthmatic mice.
abstract: Allosamidins, metabolites of Streptomyces with strong inhibitory activities toward family 18 chitinases, show a variety of biological activities in various organisms. We prepared photoaffinity and biotinylated probes of allosamidin and demethylallosamidin, the N-demethyl derivative that shows much stronger anti-asthmatic activity than allosamidin. Mild acid hydrolysis of allosamidins afforded mono-amine derivatives, which were amidated to prepare probes with a photoactivatable aryl azide and/or biotin moieties. The derivatives with an N-acyl group at C-2 of the D-allosamine residue at the non-reducing end of allosamidins inhibited Trichoderma chitinase as strongly as the original compounds. Since the target of allosamidins in asthma is unclear, photoaffinity probes were used to analyze allosamidin-binding proteins in bronchoalveolar lavage (BAL) fluid in IL-13-induced asthmatic mice. Ym1, a chitinase-like protein, was identified as the main allosamidin-binding protein among proteins whose expression was upregulated by IL-13 in BAL fluid. Binding of allosamidins with Ym1 was confirmed by the experiments with photoaffinity probes and recombinant Ym1.
authors.affiliation: Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bnkyo-ku, Japan.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Sato
authors.forename: Shigeo
authors.initials: S
authors.lastname: Suzuki
authors.forename: Seiko
authors.initials: S
authors.lastname: Muraoka
authors.forename: Naoya
authors.initials: N
authors.lastname: Kikuchi
authors.forename: Naotaka
authors.initials: N
authors.lastname: Noda
authors.forename: Takafumi
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Inoue
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Nagasawa
authors.forename: Shohei
authors.initials: S
authors.lastname: Sakuda
mesh_terms: D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use; D000818:Animals; D018927:Anti-Asthmatic Agents / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D001249:Asthma / Q000188:drug therapy* / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D002688:Chitinases / Q000037:antagonists & inhibitors*; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000627:therapeutic use; D037102:Lectins / Q000378:metabolism; D008297:Male; D051379:Mice; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D013302:Streptomyces / Q000737:chemistry; D014312:Trisaccharides / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: D018927:Anti-Asthmatic Agents; D004791:Enzyme Inhibitors; D037102:Lectins; D011994:Recombinant Proteins; D014312:Trisaccharides; C054851:allosamidin; C070537:methyl-N-demethylallosamidin; D002688:Chitinases; C429541:Chil3 protein, mouse; D001619:beta-N-Acetylhexosaminidases; D000117:Acetylglucosamine