title: Rational design, synthesis, and characterization of novel inhibitors for human beta1,4-galactosyltransferase.
abstract: An affinity labeling reagent, uridine 5'-(6-amino-{2-[(7-bromomethyl-2-naphthyl)methoxycarbonylmethoxy]ethoxy}acetyl-6-deoxy-alpha-D-galactopyranosyl) diphosphate (1a), was designed on the basis of 3D docking simulation and synthesized to investigate the functional role of Trp310 residue located in the small loop near the active site of human recombinant galactosyltransferase (betaGalT-1). Mass spectrometric analysis revealed that the Trp310 residue of betaGalT1 can be selectively modified with the naphthylmethyl group of compound 1a at the C-3 position of the indole ring. This result motivated us to synthesize novel uridine-5'-diphosphogalactose (UDP-Gal) analogues as candidates for mechanism-based inhibitors for betaGalT-1. We found that uridine 5'-(6-O-[10-(2-naphthyl)-3,6,9-trioxadecanyl]-alpha-d-galactopyranosyl) diphosphate (2) is the strongest inhibitor (K(i) = 1.86 microM) against UDP-Gal (Km = 4.91 microM) among compounds reported previously. A cold spray ionization time-of-flight mass spectrometry study demonstrated that the complex of this inhibitor and betaGalT-1 cannot interact with an acceptor substrate in the presence of Mn2+.
authors.affiliation: Division of Biological Sciences, Frontier Research Center for Post-Genome Science and Technology, Graduate School of Science, Hokkaido University, N-21, W-11, Sapporo 001-0021, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Takaya
authors.forename: Noriko
authors.initials: N
authors.lastname: Nagahori
authors.forename: Masaki
authors.initials: M
authors.lastname: Kurogochi
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Furuike
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Miura
authors.forename: Kenji
authors.initials: K
authors.lastname: Monde
authors.forename: Yuan Chuan
authors.initials: YC
authors.lastname: Lee
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D000345:Affinity Labels / Q000138:chemical synthesis* / Q000737:chemistry; D001665:Binding Sites; D015195:Drug Design; D005700:Galactosyltransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014364:Tryptophan / Q000737:chemistry; D014539:Uridine Diphosphate Sugars / Q000138:chemical synthesis* / Q000737:chemistry
chemical_list: D000345:Affinity Labels; D011994:Recombinant Proteins; D014539:Uridine Diphosphate Sugars; C505170:uridine 5'-(6-O-(10-(2-naphthyl)-3,6,9-trioxadecanyl)galactopyranosyl) diphosphate; C505169:uridine 5'-(6-amino-(2-((7-bromomethyl-2-naphthyl)methoxycarbonylmethoxy)ethoxy)acetyl-6-deoxygalactopyranosyl) diphosphate; D014364:Tryptophan; D005700:Galactosyltransferases; C493310:beta1,4-galactosyltransferase, human