title: Molecular interaction of imino sugars with human alpha-galactosidase: Insight into the mechanism of complex formation and pharmacological chaperone action in Fabry disease.
abstract: Enzyme enhancement therapy (EET) for Fabry disease involving imino sugars has been developed and attracted interest. It is thought that imino sugars act as pharmacological chaperones for wild-type and mutant alpha-galactosidases (GLAs) in cells, but the mechanisms underlying the molecular interactions between the imino sugars and the enzyme have not been clarified yet. We examined various kinds of imino sugars and found that galactostatin bisulfite (GBS) inhibited GLA in vitro and increased the enzyme activity in cultured Fabry fibroblasts as in the case of 1-deoxygalactonojirimycin (DGJ). Then, we analyzed the molecular interactions between the imino sugars and recombinant human GLA by means of isothermal titration calorimetry and surface plasmon resonance biosensor assays, and first determined the thermodynamic and binding-kinetics parameters of imino sugar and GLA complex formation. The results revealed that DGJ bound to the enzyme more strongly than GBS, the binding of DGJ to the enzyme protein being enthalpy-driven. In the case of GBS, the reaction was mainly enthalpy-driven, but there was a possibility that entropy-driven factors were involved in the binding. Structural analysis in silico revealed that both the chemicals fit into the active-site pocket and undergo hydrogen bonding with residues comprising the active-site pocket including the catalytic ones. The side chain of GBS was oriented towards the entrance of the active-site pocket, and thus it could be in contact with residues comprising the wall of the active-site pocket. Thermodynamic, kinetic and structural studies should provide us with a lot of information for improving EET for Fabry disease.
authors.affiliation: Department of Analytical Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Sugawara
authors.forename: Youichi
authors.initials: Y
authors.lastname: Tajima
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kawashima
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tsukimura
authors.forename: Seiji
authors.initials: S
authors.lastname: Saito
authors.forename: Kazuki
authors.initials: K
authors.lastname: Ohno
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Iwamoto
authors.forename: Toshihide
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Kohji
authors.initials: K
authors.lastname: Itoh
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sakuraba
mesh_terms: D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D020134:Catalytic Domain; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D000795:Fabry Disease / Q000201:enzymology* / Q000473:pathology; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D005688:Galactosamine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D050111:Imino Sugars / Q000737:chemistry / Q000494:pharmacology*; D007700:Kinetics; D008958:Models, Molecular; D013816:Thermodynamics; D000519:alpha-Galactosidase / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D050111:Imino Sugars; C051195:5-amino-5-deoxygalactopyranose; D017485:1-Deoxynojirimycin; D005688:Galactosamine; C090092:migalastat; D000519:alpha-Galactosidase