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Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human ?-galactosidase.

ABSTRACT: ?-D-Galactosidase (?-Gal) is an exoglycosidase that cleaves ?-galactosides from glycoproteins, sphingolipids and keratan sulfate. This study reports the expression, purification, crystallization and preliminary X-ray crystallographic analysis of human lysosomal ?-Gal. The sitting-drop vapour-diffusion method was used to crystallize ?-Gal in complexes with its product galactose and with the inhibitor 1-deoxygalactonojirimycin. The resulting crystals were isomorphous and belonged to space group P2(1). The crystals of the ?-Gal-galactose and the ?-Gal-inhibitor complexes had unit-cell parameters a = 94.8, b = 116.1, c = 140.3 Å, ? = 92.2° and a = 94.8, b = 116.0, c = 140.3 Å, ? = 92.2°, respectively. Diffraction data were collected to 1.8 Å resolution for both crystals.

SUBMITTER: Usui K

PROVIDER: S-EPMC3253840 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature

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Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human β-galactosidase.

Usui Kimihito K Ohto Umeharu U Ochi Toshinari T Shimizu Toshiyuki T Satow Yoshinori Y

Acta crystallographica. Section F, Structural biology and crystallization communications 20111224 Pt 1

β-D-Galactosidase (β-Gal) is an exoglycosidase that cleaves β-galactosides from glycoproteins, sphingolipids and keratan sulfate. This study reports the expression, purification, crystallization and preliminary X-ray crystallographic analysis of human lysosomal β-Gal. The sitting-drop vapour-diffusion method was used to crystallize β-Gal in complexes with its product galactose and with the inhibitor 1-deoxygalactonojirimycin. The resulting crystals were isomorphous and belonged to space group P2 ...[more]

PMID: 22232177

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