title: The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases.
abstract: alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, 01003, USA.
authors.forename: Nathaniel E
authors.initials: NE
authors.lastname: Clark
authors.forename: Scott C
authors.initials: SC
authors.lastname: Garman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D018360:Crystallography, X-Ray; D006801:Humans; D007313:Insecta; D016464:Lysosomal Storage Diseases / Q000201:enzymology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D048809:alpha-N-Acetylgalactosaminidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C482722:NAGA protein, human; D048809:alpha-N-Acetylgalactosaminidase
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