Phosphatases: Applications of new methods for their separation

Dean, P.D.G.; Willetts, S.R.; Blanch, Jaime.

doi:10.1016/0003-2697(71)90152-7

Cited by 12 publications

(1 citation statement)

References 7 publications

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“…Other affinity-chromatography steps that were tried proved unsuccessful. Dean et al (1971) used cellulose phosphate to retain phosphatases from bacterial sources, but we were unable to achieve adequate retention with the enzyme from human liver.…”

Section: Resultsmentioning

confidence: 99%

Affinity purification and some molecular properties of human liver alkaline phosphatase

Trepanier

Seargeant

Stinson

1976

Biochemical Journal

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Alkaline phosphatase from human liver was purified to homogeneity. The purification procedure included solubilization with butanol, fractionation with acetone, and chromatography on concanavalin A-Sepharose, DEAE-cellulose, Sephadex G-200 and DEAE-Sephadex. Purity was established by standard and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The isoelectric point of the protein was determined to be 4.0. Sephadex-gel filtration gave a mol.wt. of 146000, although a higher value was obtained in the presence of 100mM-NaC1. The subunit mol.wt. 76700, was determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Neuraminidase treatment resulted in two enzyme-activity bands on isoelectric-focused gels with isoelectric points of 6.6 and 6.8. The desialylated enzyme gave only one protein band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis with a subunit molecular weight indistinguishable from that of the non-neuraminidase-treated protein. The desialylated enzyme was more readily denatured by sodium dodecyl sulphate in the presence of mercaptoethanol than was the native enzyme.

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Section: Resultsmentioning

confidence: 99%

Affinity purification and some molecular properties of human liver alkaline phosphatase

Trepanier

Seargeant

Stinson

1976

Biochemical Journal

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Träger‐gebundene, biologisch aktive Substanzen und ihre Anwendung

Orth¹,

Brümmer²

1972

Angewandte Chemie

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Durch Bindung von Enzymen an geeignete Träger lassen sich stabile Katalysatoren gewinnen. Es werden die wichtigsten Methoden ihrer Herstellung sowie ihre Eigenschaften erläutert. Besonders interessant erscheint die Anwendung träger‐gebundener Enzyme in der biochemischen Routineanalytik als „unlösliche Reagentien”︁ für Analysenautomaten und für Enzymelektroden. Für präparative Stoffumwandlungen und Synthesen bieten die träger‐gebundenen Enzyme den Vorteil, die bisher meist diskontinuierlichen Prozesse in kontinuierliche umwandeln und proteinfreie Reaktionsprodukte erhalten zu können. Durch Verwendung träger‐gebundener Proteasen als spezifische Adsorbentien läßt sich eine Reihe natürlich vorkommender Inhibitoren in einfacher Weise isolieren. Dieses Prinzip, das der Affinitätschromatographie zugrunde liegt, wird heute umgekehrt auch zur Reinigung von Enzymen und anderen biologisch aktiven Substanzen an entsprechenden träger‐gebundenen Effektoren ausgenutzt.

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Carrier‐Bound Biologically Active Substances and Their Applications

Orth

Brümmer

1972

Angew. Chem. Int. Ed. Engl.

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Phosphatases: Applications of new methods for their separation

Cited by 12 publications

References 7 publications

Affinity purification and some molecular properties of human liver alkaline phosphatase

Affinity purification and some molecular properties of human liver alkaline phosphatase

Träger‐gebundene, biologisch aktive Substanzen und ihre Anwendung

Carrier‐Bound Biologically Active Substances and Their Applications

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