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

E-ISSN： 1744-3091|68|1|73-77

ISSN： 1744-3091

Source： Acta Crystallographica Section F, Vol.68, Iss.1, 2012-01, pp. : 73-77

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Abstract

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 P2₁. The crystals of the β‐Gal–galactose and the β‐Gal–inhibitor complexes had unit‐cell parameters a = 94.8, b = 116.1, c = 140.3 Å, β = 92.2° and a = 94.8, b = 116.0, c = 140.3 Å, β = 92.2°, respectively. Diffraction data were collected to 1.8 Å resolution for both crystals.