PMID: 41823Sep 1, 1979

A quantitative histochemical technique for the characterisation of alpha-glucosidases in the brush-border membrane of rat jejunum

Histochemistry
S GutschmidtE O Riecken

Abstract

A quantitative histochemical method to determine the Km and Vmax of alpha-glucosidases in the intestinal epithelium without disruption of the cellular structure is described. 2-Naphthyl-alpha-D-glucoside was used as substrate and hexazonium-p-rosaniline as coupling agent. Using a Leitz MPV2 microdensitometer and a field measuring 4 X 4 micrometers, and reading the test samples against a blank focused on the lamina propria, we observed that the intensity of the colour was a linear function of both the incubation time up to 20 min, and the thickness of the slice up to 20 micrometers. The ratio between the extinction at the absorption maximum and at a second wave-length was constant, whatever the intensity of the colour. By determining the relationship between the extinction and the substrate concentration under standard conditions (slice thickness of of 10 micrometers and incubation time of 10 min), we obtained a saturation curve described by a Km of 0.68 +/- 0.038 mM and a Vmax of 1.41 +/- 0.039 A lambda 480 . 10(-2) . micrometers-1 . min-1. When the hydrolysis of the same substrate by a homogenate of jejunal mucosa was examined biochemically under comparable conditions, a Km of 0.64 +/- 0.012 mM and a Vmax of 57.3 +/- 0.70 mU/m...Continue Reading

References

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Citations

Jan 1, 1981·Histochemistry·S Gutschmidt, R Gossrau
Jan 1, 1982·Archives of Dermatological Research·S Gutschmidt, D Tsambaos
Jan 1, 1983·Research in Experimental Medicine. Zeitschrift Für Die Gesamte Experimentelle Medizin Einschliesslich Experimenteller Chirurgie·S GutschmidtE O Riecken
Jan 1, 1980·Research in Experimental Medicine. Zeitschrift Für Die Gesamte Experimentelle Medizin Einschliesslich Experimenteller Chirurgie·J W RobinsonV Mirkovitch
Jan 23, 1990·Biochimica Et Biophysica Acta·M W SmithA J Collins
Jul 10, 1991·Biochimica Et Biophysica Acta·M W SmithM A Peacock
Jan 1, 1984·Comparative Biochemistry and Physiology. A, Comparative Physiology·D CremaschiM W Smith
Jan 1, 1984·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·M W Smith, R M Moor

Related Concepts

Lysosomal alpha-Glucosidase
Plasma Membrane
Densitometry
Glucosidase
Cytochemistry
Hydrogen-Ion Concentration
Structure of Intestinal Gland
Jejunum
Microvilli
Substrate Specificity

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