Heterogeneity of Beta-Galactosidase from Rabbit Spleen

Fj, Rodriguez-Berrocal; Mn, Pérez-Gonzalez; Ja, Cabezas

doi:10.1159/000469574

Cited by 8 publications

(1 citation statement)

References 12 publications

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“…Some of the several minor bands could represent the "protective factor" and "neuraminidase subunit" proteins, which have been reported to copurify with acid ß-galactosidase from other species (15,16). The presence of other enzymes was, however not examined, but should have been revealed m the haemagglutination tests.…”

Section: Discussionmentioning

confidence: 99%

Isolation of a β-Galactosidase from Chicken Liver

Javeri¹,

Uhlenbruck²

1984

Clinical Chemistry and Laboratory Medicine

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Summary:In search of a ß-galactosidase which specifically hydrolyses 1-»3 bound galactose residues in galacto-glycoconjugates, an acid ß-galactosidase from chicken liver was investigated. The Isolation procedure involved ammonium sulphate precipitation followed by lectin chromatography on Con A-Sepharose 4B, ionexchange chromatography on DEAE-cellulose, gel filtration on Sepliarose 6B and affinity chromatography on/7-aminophenyl-thio-ß-D-galactoside-agarose. The ß-galactosidase was purified 3000-fold with 11% recovery of enzyme activity. The purified protein showed an apparent molecular mass of above 200000 in SDSpolyacrylamide gel electrophoresis. A few minor bands were also present. The reduced and denatured ß-galactosidase migrated äs a single major band with an apparent molecular mass of 67000. The enzyme released galactose from lactose and from the synthetic Substrates Galßl->3Gal, Galßl-»6Gal and Galßl-»3Ara. However, the enzyme did not release galactose from the snail gland galactans and the high molecular weight galacto-glycoconjugates and it did not hydrolyse the peanut agglutinin receptor of the red blood cell membrane. Isolierung einer ß-Galaktosidase aus HühnerleberZusammenfassung: Auf der Suche nach ß-Galaktosidasen, die spezifisch ßl->3 gebundene Galaktosereste hydrolysieren, wie z.B. in verschiedenen Galakto-Glykokonjugaten, wurde eine saure ß-Galaktosidase aus Hühnerleber untersucht. Das Isolierungsverfahren beinhaltete Ammoniumsulfat-Fällung mit anschließender Lektin-Chrömatographie an Con A-Sepharose 4B, lonenaustausch-Chromatographie an DEAE-Cellulose, Gelfiltration an Sepharose 6B und Affinitätschromatographie an p-Aminophenyl-thio-ß-D-galaktosid-Agarose. Die ß-Galaktosidase wurde 3000fach angereichert mit einer verbleibenden Enzymaktivität von 11%. Das gereinigte Protein zeigte in der SDS-Polyacrylamid-Gelelektrophorese ein Molekulargewicht von 200000 Dalton. Zusätzlich traten einige kleinere Banden auf. Nach Reduktion und Denaturierung wanderte die ß-Galaktosidase als eine einheitliche Hauptbande mit einem Molekulargewicht von 67000 Dalton. Das Enzym hydrolysierte Galaktose aus Lactose und aus den synthetisierten Substraten Galßl-*3Gal, Galßl-> 6Gal und Galßl-»3Ara. Die Spaltung von Galaktose aus dem Galaktan der Schneckeneiweißdrüse und aus anderen hochmolekularen Galakto-Glykokorijugaten war allerdings nicht möglich, und auch der Erdnußag-glutinin-Rezeptor der Erythrocytenmembran ließ sich durch das Enzym nicht inaktivieren. Intrödücfiongalacto-glycoconjugates (e.g. the so-called peanut ß-Galactosidases (EC 3.2.1.23) occur ubiquitously receptor (PNA) or lectins specific for N-acetyl-lacin animals, plants and microorganisms (1). They tosamine) has beconie of great importance in imhavegainedincreasinginterest,beeausetheinactivamunochemistry, especially with respect to such retion of ß-D-galactosidic lectin receptors in different ceptors in tumour cells (2).

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Section: Discussionmentioning

confidence: 99%