Aggregation-dissociation and stability of acid β-galactosidase purified from porcine spleen

Yamamoto, Yoshimi; Nishimura, Kenji

doi:10.1016/0020-711x(86)90038-8

Cited by 6 publications

(3 citation statements)

References 22 publications

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“…Dissociation constants at some protein complex have been found to be pressure-sensitive [19]. However, other authors have also reported the existence of several forms of lysosomal fl-galactosidase differing in molecular size [20][21][22][23].…”

Section: D56mentioning

confidence: 99%

“…The molecular masses reported previously for the NGCcomplex or ,-galactosidase multimeric forms in various tissues were usually obtained by gel filtration using standard proteins of known molecular mass to calibrate the column [20][21][22][23]. These values must be considered as approximate since we found that the NGC-complex is highly hydrated and/or asymmetric, a situation which could lead to relatively large systematic errors in molecular mass determinations by gel filtration, which assumes that all proteins behave as a non-hydrated sphere [24].…”

Section: D56mentioning

confidence: 99%

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Structure of the lysosomal neuraminidase–β-galactosidase–carboxypeptidase multienzymic complex

Potier

Michaud

Tranchemontagne

et al. 1990

Biochemical Journal

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Lysosomal neuraminidase (sialidase; EC 3.2.1.18) and beta-galactosidase (EC 3.2.1.23), together with a carboxypeptidase, the so-called 'protective protein', were co-purified from the human placenta by affinity chromatography on a concanavalin A-Sepharose column followed by a thiogalactoside-agarose affinity column for beta-galactosidase. Analysis of the purified material by gel-filtration h.p.l.c. revealed three distinct molecular forms, all with high beta-galactosidase specific activity, but only the largest one expressed neuraminidase activity. Rechromatography of each individual species separately indicated that all three are in fact part of an equilibrium system (the neuraminidase-beta-galactosidase-carboxypeptidase complex or NGC-complex) and that these species undergo slow conversion into one another through dissociation and association of protomeric components. Each species was sufficiently stable for the determination of their hydrodynamic properties by gel-filtration h.p.l.c. and sedimentation velocity. The largest species had an apparent sedimentation coefficient S20.w, of 18.8 S and a Stokes' radius of 8.5 nm, giving a molecular mass of 679 kDa and a fractional ratio, f/f min, of 1.47. The latter value indicates that the macromolecule is asymmetric or highly hydrated. This large species is composed of four types of polypeptide chains of molecular mass 66 kDa (neuraminidase), 63 kDa (beta-galactosidase), 32 kDa and 20 kDa (carboxypeptidase heterodimer). The 32 kDa and 20 kDa protomers are linked together by a disulphide bridge. Glycopeptidase F digestion of the NGC-complex transformed the diffuse 66-63 kDa band on the SDS gel into two close but sharp bands at 58 and 56 kDa. The two smaller species which were separated on the h.p.l.c. column correspond to tetrameric and dimeric forms of the 66-63 kDa protomers and express exclusively beta-galactosidase activity. Treatment of the NGC-complex with increasing concentrations of guanidinium hydrochloride up to 1.5 M also resulted in dissociation of the complex into the same smaller species mentioned above plus two protomers of molecular mass around 60 and 50 kDa. A model of the largest molecular species as a hexamer of the 66-63 kDa protomers associated to five carboxypeptidase heterodimers (32 kDa and 20 kDa) is proposed

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Section: D56mentioning

confidence: 99%

Section: D56mentioning

confidence: 99%

Structure of the lysosomal neuraminidase–β-galactosidase–carboxypeptidase multienzymic complex

Potier

Michaud

Tranchemontagne

et al. 1990

Biochemical Journal

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“…In multicellular organisms, β-galactosidases are located in the lysosomes or in the cytosol, depending on their function and substrate specificity, acting at acidic or neutral pH-optima. A change in the pH-environment has been found to play an important role in protein folding and, therefore, influences the activity properties [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

Identification, Characterization, and Expression of a β-Galactosidase from Arion Species (Mollusca)

et al. 2022

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β-Galactosidases (β-Gal, EC 3.2.1.23) catalyze the cleavage of terminal non-reducing β-D-galactose residues or transglycosylation reactions yielding galacto-oligosaccharides. In this study, we present the isolation and characterization of a β-galactosidase from Arion lusitanicus, and based on this, the cloning and expression of a putative β-galactosidase from Arion vulgaris (A0A0B7AQJ9) in Sf9 cells. The entire gene codes for a protein consisting of 661 amino acids, comprising a putative signal peptide and an active domain. Specificity studies show exo- and endo-cleavage activity for galactose β1,4-linkages. Both enzymes, the recombinant from A. vulgaris and the native from A. lusitanicus, display similar biochemical parameters. Both β-galactosidases are most active in acidic environments ranging from pH 3.5 to 4.5, and do not depend on metal ions. The ideal reaction temperature is 50 °C. Long-term storage is possible at up to +4 °C for the A. vulgaris enzyme, and at up to +20 °C for the A. lusitanicus enzyme. This is the first report of the expression and characterization of a mollusk exoglycosidase.

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Inherited lysosomal storage disease associated with deficiencies of β‐galactosidase and α‐neuraminidase in sheep

et al. 1988

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Histopathologic, ultrastructural and Golgi impregnation studies disclosed lesions characteristic of a neuronal lysosomal storage disease in related sheep with onset of neurologic signs at 4-6 months. Biochemical and enzymatic evaluation disclosed storage of GM1 ganglioside, asialo-GM1, and neutral long chain oligosaccharides in brain, urinary excretion of neutral long chain oligosaccharides, and deficiencies of lysosomal beta-galactosidase and alpha-neuraminidase. Retrospective and limited prospective genetic studies suggested autosomal recessive inheritance. A gene-dosage effect on beta-galactosidase levels was documented in fibroblasts from putative heterozygous sheep. Fibroblasts from affected sheep did not have increased beta-galactosidase activity after incubation with the protease inhibitor, leupeptin. In some aspects this disease is similar to GM1 gangliosidosis, but is unique in that a genetic defect in lysosomal beta-galactosidase may cause the deficiency of lysosomal alpha-neuraminidase.

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Aggregation-dissociation and stability of acid β-galactosidase purified from porcine spleen

Cited by 6 publications

References 22 publications

Structure of the lysosomal neuraminidase–β-galactosidase–carboxypeptidase multienzymic complex

Structure of the lysosomal neuraminidase–β-galactosidase–carboxypeptidase multienzymic complex

Identification, Characterization, and Expression of a β-Galactosidase from Arion Species (Mollusca)

Inherited lysosomal storage disease associated with deficiencies of β‐galactosidase and α‐neuraminidase in sheep

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