The electrophoretic separation of human β-galactosidases on cellulose acetate

Fluharty, Arvan L.; Lassila, Elton L.; Porter, Myna T.; Kihara, Hayato

doi:10.1016/0006-2944(71)90083-4

Cited by 22 publications

(4 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This suggests that the residual p-galactosidases in the brain of patients with these two types of Gm, gangliosidosis are actually different. Previous studies in human brain and rabbit brain [5, 61 revealed three p-galactosidase isozymes, two of which migrate towards the anode; one does not migrate from the origin and is thought to be particlebound [3]. I n our study, two p-galactosidase bands were preseni on both cellufose acetate and starch gel after electrophoresis.…”

supporting

confidence: 55%

Brain β-Galactosidase and Gm1 Gangliosidosis

1974

View full text Add to dashboard Cite

supporting

confidence: 55%

Brain β-Galactosidase and Gm1 Gangliosidosis

1974

View full text Add to dashboard Cite

“…Electrophoresis of tissue supernatant (l0op1; 50%, w/v) was carried out on Cellogel (Whatman Lab Sales, Maidstone, Kent, U.K.) at pH 6.5 by the method of Fluharty et al (1971) for 2h at 4°C using a potential of 160V and a current of 2.5mA/band. Acidic and neutral a-mannosidase were detected after electrophoresis by placing Whatman 3MM paper soaked in 2 mM-methylumbelliferyl a-D-mannopyranoside at pH4.0 or pH6.5 respectively on the gel for 30min at 37°C.…”

Section: Electrophoresis On Cellulose Acetatementioning

confidence: 99%

Characterization of the mutant α-mannosidase in bovine mannosidosis

Burditt

Phillips

Robinson

et al. 1978

Biochemical Journal

View full text Add to dashboard Cite

Residual acidic alpha-mannosidase, varying in amount up to approx. 15% of normal values, can be measured in various organs of a calf with mannosidosis. The highest specific activity and relative proportion of residual activity were found in the liver. Chromatography on DEAE-cellulose showed that the residual activity was associated with two components, which were eluted at comparable positions with those found in normal tissues. The residual activity had a lower thermal stability and a higher K(m) value for a synthetic substrate than did the normal enzyme. No differences in molecular weight or electrophoretic mobility between normal acidic alpha-mannosidase and the residual activity were observed by gel filtration and electrophoresis on cellulose acetate respectively. The isoelectric focusing profiles for the alpha-mannosidase in the normal and pathological livers were very similar. It is suggested that a mutant enzyme, resulting from a mutation in a structural gene, accounts for the residual acidic alpha-mannosidase in mannosidosis. The mutant enzyme, which cross-reacts with antiserum raised against normal bovine acidic alpha-mannosidase, is present at a decreased concentration compared with the normal enzyme. There is a correlation between the concentrations of residual activity and cross-reacting material in mannosidosis. alpha-Mannosidase with a pH optimum of 5.75 and which is activated by Zn(2+) was also detected in the liver of the calf with mannosidosis. However, it is probably not a product of the defective gene because addition of Zn(2+) indicated that it was also present in normal tissues.

show abstract

“…Flectrophoresis. Cellogel electrophoresis was carried out as described by Fluharty et al (1971), with modification, for 2h at 4°C and 3mA/cm. The buffer was 0.04M-potassium phosphate, pH6.6.…”

Section: Molecular Weight Det4rminationmentioning

confidence: 99%

Isolation and properties of α-D-mannosidase from human kidney

Marinkovic

1976

Biochemical Journal

View full text Add to dashboard Cite

Alpha-D-Mannosidase activity exists in three forms that can be separated by DEAE-cellulose chromatography, alpha-D-Mannosidase was isolated from human kidney in a homogeneous state, and was purified 2100-fold, with p-nitrophenyl alpha-D-mannoside as substrate. The purified alpha-D-mannosidase was practically free from all other glycosidases tested. The Km of the synthetic substrate with the enzyme was 1 X 10(-3) M and the pH optimum 4.5. It was inhibited by heavy metals, sodium dodecyl sulphate, urea and compounds that react with the thiol groups, and was activated by Zn2+, Na+, 2-mercaptoethanol, human albumin and gamma-globulin. The mol. wt. of the enzyme was estimated to be 180 000 +/- 4500. After pretreatment with 2-mercaptoethanol and sodium dodecyl sulphate, alpha-D-mannosidase dissociated into subunits of mol. wts. of 58 000 +/- 600 and 30 000 +/- 380 respectively. Subunits of the same molecular weights were also obtained after the enzyme was heated at 100 degrees C.

show abstract

The electrophoretic separation of human β-galactosidases on cellulose acetate

Cited by 22 publications

References 14 publications

Brain β-Galactosidase and Gm1 Gangliosidosis

Brain β-Galactosidase and Gm1 Gangliosidosis

Characterization of the mutant α-mannosidase in bovine mannosidosis

Isolation and properties of α-D-mannosidase from human kidney

Contact Info

Product

Resources

About