The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases.
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.
Clark
Nathaniel E
NE
Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, 01003, USA.
Garman
Scott C
SC
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
C482722:NAGA protein, human; D048809:alpha-N-Acetylgalactosaminidase
Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
15299374
Biochem Cell Biol. 2008 Apr;86(2):169-77
18443630
Lancet. 1987 Oct 3;2(8562):804
2889023
Structure. 2002 Mar;10(3):425-34
12005440
J Inherit Metab Dis. 1994;17(6):724-31
7707696
J Mol Biol. 2004 Mar 19;337(2):319-35
15003450
Eur J Hum Genet. 2001 Feb;9(2):91-6
11313741
J Inherit Metab Dis. 1988;11(4):349-57
3149698
N Engl J Med. 1989 Jun 29;320(26):1735-40
2733734
Protein Expr Purif. 2004 Feb;33(2):274-87
14711516
J Mol Biol. 2004 May 28;339(2):413-22
15136043
Nat Biotechnol. 2007 Apr;25(4):454-64
17401360
Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
7984417
Br J Haematol. 2008 Jan;140(1):3-12
17970801
J Hum Genet. 2004;49(1):1-8
14685826
Biochim Biophys Acta. 1977 May 12;482(1):138-44
405044
J Lipid Res. 2002 Jul;43(7):1096-104
12091494
Carbohydr Res. 2000 Nov 17;329(3):539-47
11128583
Biochem Biophys Res Commun. 1977 Aug 22;77(4):1411-7
901541
J Biol Chem. 2003 May 30;278(22):20313-8
12657636
Br J Dermatol. 2001 Feb;144(2):363-8
11251574
Nat Biotechnol. 2005 May;23(5):567-75
15877075
Methods Enzymol. 1997;276:307-26
27754618
Life Sci. 2005 Mar 4;76(16):1817-24
15698859
Structure. 1997 Dec 15;5(12):1557-69
9438862
Nucleic Acids Res. 2009 Jan;37(Database issue):D233-8
18838391
J Inherit Metab Dis. 2007 Feb;30(1):108
17171432
Cancer Lett. 2000 Sep 29;158(1):61-4
10940510
J Dermatol Sci. 2005 Jan;37(1):15-20
15619430
Biochem J. 1999 Apr 1;339 ( Pt 1):43-53
10085226
J Clin Invest. 1994 Aug;94(2):839-45
8040340
J Biol Chem. 1990 Dec 15;265(35):21859-66
2174888
Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9
2025413
J Clin Invest. 1990 Nov;86(5):1752-6
2243144
Acta Crystallogr D Biol Crystallogr. 2004 Jun;60(Pt 6):1048-56
15159564
J Med Genet. 1996 Jun;33(6):458-64
8782044
Glycobiology. 2000 Mar;10(3):251-61
10704524
Nat Rev Genet. 2002 Dec;3(12):954-66
12459725