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Histochemistry

, Volume 52, Issue 2, pp 187–197 | Cite as

Mikrochemische Untersuchung der α-d-Glucosidasen mit 4-Methylumbelliferyl-und 2-Naphthyl-α-d-glucosid

  • R. Gossrau
Article
  • 16 Downloads

Zusammenfassung

In Rohhomogenaten aus gefriergetrockneten Kryostat-schnitten von verschiedenen Rattenorganen werden die Km und Vmax der neutralen und sauren α-d-Glucosidase bestimmt und der Einfluß von pH, Substrat- und Enzymkonzentration und Inkubationszeit auf die Aktivität fluorometrisch mit 4-Methylumbelliferyl-und 2-Naphthyl-α-d-glucosid als Substraten ermittelt.

Mit den biochemischen Daten werden 2 mikrochemische Ansätze zur fluorometrischen Messung dieser Glykosidasen entwickelt und die saure und neutrale α-Glucosidase in Gruppen von Epithelzellen nach Isolierung aus gefriergetrockneten Kryostatschnitten von Nebenhoden, Jejunum, Ilium, Niere und Leber untersucht. Im Vergleich zum 2-Naphthylderivat sind beide α-Glucosidasen mit 4-Methylumbelliferyl-α-d-glucosid weniger aktiv. Allerdings fluoresziert 4-Methylumbelliferon etwa 100mal intensiver als 2-Naphthol, so daß das Methylumbelliferonderivat zur Messung der α-Glucosidasen speziell in schwach aktiven Zellen der 2-Naphthylverbindung vorzuziehen ist.

Microchemical investigation of α-d-glucosidases using 4-methylumbelliferyl-and 2-naphthyl-α-d-glucoside

Summary

In crude homogenates prepared from freeze-dried cryostate sections of various rat organs the Km and Vmax of acid and neutral α-glucosidase as well as the effect of the pH, substrate and enzyme concentration and the incubation time on the activity were determined fluorometrically with 4-methylumbelliferyl-and 2-naphthyl α-d-glucoside as substrates.

On the basis of the biochemical data 2 assays were developed for the microchemical measurement of both α-glucosidases in groups of epithelial cells isolated from freeze-dried cryostate sections of the epididymis, jejunum, ilium, liver and kidney of suckling and adult rats. The rate of hydrolysis of 2-naphthyl and 4-methylumbelliferyl α-d-glucoside differs moderately. However, due to the higher sensitivity of 4-methylumbelliferone the methylumbelliferyl derivative is preferable especially for the evaluation of α-d-glucosidases in cells with low enzyme activity.

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Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • R. Gossrau
    • 1
  1. 1.Anatomisches Institut der Universität WürzburgWürzburgGermany

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