title: Structural basis of pharmacological chaperoning for human β-galactosidase.
abstract: GM1 gangliosidosis and Morquio B disease are autosomal recessive diseases caused by the defect in the lysosomal β-galactosidase (β-Gal), frequently related to misfolding and subsequent endoplasmic reticulum-associated degradation. Pharmacological chaperone (PC) therapy is a newly developed molecular therapeutic approach by using small molecule ligands of the mutant enzyme that are able to promote the correct folding and prevent endoplasmic reticulum-associated degradation and promote trafficking to the lysosome. In this report, we describe the enzymological properties of purified recombinant human β-Gal(WT) and two representative mutations in GM1 gangliosidosis Japanese patients, β-Gal(R201C) and β-Gal(I51T). We have also evaluated the PC effect of two competitive inhibitors of β-Gal. Moreover, we provide a detailed atomic view of the recognition mechanism of these compounds in comparison with two structurally related analogues. All compounds bind to the active site of β-Gal with the sugar-mimicking moiety making hydrogen bonds to active site residues. Moreover, the binding affinity, the enzyme selectivity, and the PC potential are strongly affected by the mono- or bicyclic structure of the core as well as the orientation, nature, and length of the exocyclic substituent. These results provide understanding on the mechanism of action of β-Gal selective chaperoning by newly developed PC compounds.
authors.affiliation: From the Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Hironori
authors.initials: H
authors.lastname: Suzuki
authors.affiliation: From the Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Umeharu
authors.initials: U
authors.lastname: Ohto
authors.affiliation: the Division of Functional Genomics, Research Center for Bioscience and Technology, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan.
authors.forename: Katsumi
authors.initials: K
authors.lastname: Higaki
authors.affiliation: the Department of Organic Chemistry, Faculty of Chemistry, University of Seville, Profesor García González 1, E-41012 Seville, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Mena-Barragán
authors.affiliation: the Department of Organic Chemistry, Faculty of Chemistry, University of Seville, Profesor García González 1, E-41012 Seville, Spain.
authors.forename: Matilde
authors.initials: M
authors.lastname: Aguilar-Moncayo
authors.affiliation: the Department of Organic Chemistry, Faculty of Chemistry, University of Seville, Profesor García González 1, E-41012 Seville, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Ortiz Mellet
authors.affiliation: the Division of Functional Genomics, Research Center for Bioscience and Technology, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Nanba
authors.affiliation: the Institute for Chemical Research (IIQ), CSIC, University of Sevilla, Americo Vespucio 49, Isla de la Cartuja, E-41092 Sevilla, Spain.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Garcia Fernandez
authors.affiliation: the International University of Health and Welfare Graduate School, Kita Kanemaru, Otawara, Tochigi 324-8501, Japan, and.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: From the Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan, CREST, Japan Science and Technology Agency, 4-1-8 Honcho Kawaguchi, Saitama 332-0012, Japan shimizu@mol.f.u-tokyo.ac.jp.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Shimizu
mesh_terms: D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D053138:Cyclohexenes / Q000737:chemistry / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004795:Enzyme Stability / Q000187:drug effects; D016537:Gangliosidosis, GM1 / Q000201:enzymology* / Q000235:genetics; D006595:Hexosamines / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D050111:Imino Sugars / Q000737:chemistry / Q000494:pharmacology; D007294:Inositol / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D007700:Kinetics; D008958:Models, Molecular; D015394:Molecular Structure; D009085:Mucopolysaccharidosis IV / Q000201:enzymology* / Q000235:genetics; D009154:Mutation; D017434:Protein Structure, Tertiary; D055672:Static Electricity; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics
chemical_list: D053138:Cyclohexenes; D004791:Enzyme Inhibitors; D006595:Hexosamines; D050111:Imino Sugars; C116532:N-octyl-beta-valienamine; C483225:proto-quercitol; D017485:1-Deoxynojirimycin; C030791:valienamine; D007294:Inositol; D001616:beta-Galactosidase; C013626:nojirimycin
keywords: Crystal Structure; Galactose; Glycosidases; Lysosomal Storage Disease; X-ray Crystallography
references.citation: Expert Opin Ther Pat. 2011 Jun;21(6):885-903
references.pmid: 21457079
references.citation: Hum Mutat. 2009 Aug;30(8):1214-21
references.pmid: 19472408
references.citation: Org Biomol Chem. 2011 May 21;9(10):3698-713
references.pmid: 21451818
references.citation: Carbohydr Res. 2013 Mar 7;368:8-15
references.pmid: 23314299
references.citation: Mol Ther. 2013 Mar;21(3):526-32
references.pmid: 23337983
references.citation: EMBO Mol Med. 2009 Aug;1(5):268-79
references.pmid: 20049730
references.citation: J Biol Chem. 1973 Oct 10;248(19):6772-80
references.pmid: 4270451
references.citation: Biol Chem. 2008 Jan;389(1):1-11
references.pmid: 18095864
references.citation: Acta Paediatr. 2008 Apr;97(457):88-93
references.pmid: 18339196
references.citation: Chem Biol. 2011 Dec 23;18(12):1521-6
references.pmid: 22195554
references.citation: Annu Rev Genomics Hum Genet. 2012;13:307-35
references.pmid: 22970722
references.citation: Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
references.pmid: 15299374
references.citation: Anal Biochem. 1972 Jun;47(2):527-38
references.pmid: 5036460
references.citation: Am J Hum Genet. 1991 Nov;49(5):1091-3
references.pmid: 1928092
references.citation: Chemistry. 2012 Jul 2;18(27):8527-39
references.pmid: 22674827
references.citation: Clin Biochem. 1999 Apr;32(3):167-70
references.pmid: 10383075
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references.pmid: 15299926
references.citation: Chembiochem. 2008 Nov 3;9(16):2612-8
references.pmid: 18833549
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Am J Hum Genet. 1991 Aug;49(2):435-42
references.pmid: 1907800
references.citation: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Jan 1;68(Pt 1):73-7
references.pmid: 22232177
references.citation: Annu Rev Med. 2006;57:283-96
references.pmid: 16409150
references.citation: Hum Mutat. 2011 Jul;32(7):843-52
references.pmid: 21520340
references.citation: Gastroenterology. 1969 May;56(5):985-6
references.pmid: 5782307
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):386-94
references.pmid: 21460457
references.citation: Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15428-33
references.pmid: 12434014
references.citation: Perspect Medicin Chem. 2009 May 26;3:7-19
references.pmid: 19812739
references.citation: Chembiochem. 2009 Nov 23;10(17):2780-92
references.pmid: 19830760
references.citation: Chem Commun (Camb). 2012 Jul 4;48(52):6514-6
references.pmid: 22618082
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Bioorg Med Chem. 2002 Jun;10(6):1967-72
references.pmid: 11937356
references.citation: Biochim Biophys Acta. 1999 Oct 8;1455(2-3):85-103
references.pmid: 10571006
references.citation: Brain Dev. 2013 Jun;35(6):515-23
references.pmid: 23290321
references.citation: Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15912-7
references.pmid: 14676316
references.citation: J Org Chem. 2000 Jan 14;65(1):136-43
references.pmid: 10813907
references.citation: J Biol Chem. 2012 Jan 13;287(3):1801-12
references.pmid: 22128166