Synopsis
Whele homogenate of guinea-pig small intestine mucosa was analysed by centrifugation in a zonal rotor. The results indicate that FAD-linked α-glycerophosphate dehydrogenase is localized in the mitochondria and that NAD-linked α-glycerophosphate dehydrogenase is a soluble phase enzyme. An enzyme hydrolysing α-naphthyl palmitate at an acid pH was localized in the lysosomes and was activated by 0.1% Triton X-roo and by freezing and thawing. An alkaline hydrolase acting on β-naphthyl laurate was localized in the ‘microsomes’. The possibility of this enzyme being different from α-naphthyl acetate hydrolase is discussed.
Similar content being viewed by others
References
Brindley, D. N. (1974). The intracellular phase of fat absorption. In:Biomembranes, Vol. 4B (ed. D. H. Smith), Ch. 12, pp. 621–71. London: Plenum Press.
Beaufay, M., Amar-Costesec, A., Thine-Sempoux, D., Wibo, M., Robbi, M. &Berthet, J. (1974). Analytical study of microsomes and isolated subcellular membranes from rat liver.J. Cell Biol. 61, 213–31.
Connock, M. J. (1973). Intestinal peroxisomes in the goldfish (Carassius auratus).Comp. Biochem. Physiol. 45A, 945–51.
Connock, M. J., Elkin, A. &Pover, W. F. R. (1971). The preparation of brush borders from the epithelial cells of the guinea-pig small intestine by zonal centrifugation.Histochem. J. 3, 11–22.
Connock, M. J. &Kirk, P. R. (1973). Identification of peroxisomes in the epithelial cells of the small intestine of the guinea-pig.J. Histochem. Cytochem. 21, 502–3.
Connock, M. J., Kirk, P. R. &Sturdee, A. P. (1974). A zonal rotor method for the preparation of microperoxisomes from epithelial cells of guinea-pig small intestine.J. Cell Biol. 61, 123–33.
Connock, M. J. &Pover, W. F. P. (1970). Catalase particles in the epithelial cells of the guinea-pig small intestine.Histochem. J. 2, 371–80.
De Duve, C., Pressman, B. C., Gianetto, R., Wattiaux, R. &Applemans, F. (1955). Tissue Fractionation Studies. 6. Intracellular distribution patterns of enzymes in rat liver tissue.Biochem. J. 60, 604–17.
Dinella, R. R., Meng, H. C. &Park, C. R. (1960). Properties of intestinal lipase.J. biol. Chem. 235 3076–81.
Gardner, R. S. (1974). A sensitive colorimetric assay for mitochondrial α-glycerophosphate dehydrogenase.Anal. Biochem.,59, 272–6.
Gee, R., McGroarty, E., Hsich, B., Wied, D. M. &Tolbert, N. E. (1974). Glycerol phosphate dehydrogenase in mammalian peroxisomes.Arch. Biochem. Biophys. 161, 187–93.
Hubscher, G., West, G. R., &Brindley, D. N. (1965). Studies on the fractionation of mucosal homogenates from the small intestine.Biochem. J. 97, 629–42.
Lojda, Z. (1974). Cytochemistry of enterocytes and of other cells in the mucous membrane of the small intestine. In:Biomembranes, Vol. 4A (ed. D. H. Smyth), p. 43. London: Plenum Press.
Mahadevan, S. &Tappel, A. L. (1968). Lysosomal lipases of rat liver and kidney.J. biol. Chem. 243, 2849–54.
Michell, R. H., Karnovsky, M. J. &Karnovsky, M. L. (1970). The distribution of some granule-associated enzymes in guinea-pig polymorphonuclear leucocytes.Biochem. J. 116, 207–16.
Novikoff, P. M. &Novikoff, A. B. (1972). Peroxisomes in absorptive cells of mammalian small intestine.J. Cell Biol. 53, 532–60.
Novikoff, A. B., Novikoff, P. M., Davis, C. &Quintana, N. (1973). Studies on microperoxisomes. V. Are microperoxisomes ubiquitous in mammalian cells?J. Histochem. Cytochem. 21, 737–55.
Pearse, A. G. E. &Riecken, E. O. (1967). Histology and cytochemistry of cells of small intestine.Brit. Med. Bull. 23, 217–22.
Peters, T. J. (1972). Subcellular fractionation of the enterocyte with special reference to peptide hydrolases. In:Peptide transport in Bacteria and Mammalian cells. Ciba Foundation Symposium, p. 107. Amsterdam: Associated Scientific Publishers.
Ope, J. L., McPherson, J. C. &Tidwell, H. C. (1966). A study of a monoglyceride-hydrolysing enzyme of intestinal mucosa.J. biol. Chem.,241, 2306–10.
Porteous, J. W. (1972). The isolation of brush borders (microvilli) from the epithelial cells of mammalian intestine. In:Subcellular Components (ed. G. D. Birnie). London: Butterworths.
Prospero, T. D. (1974). A simplified assay for succinare dehydrogenase. In:Methodological Developments in Biochemistry, Vol. 4Subcellular Studies (ed. E. Reid), pp. 411–12. London: Longman.
Ravin, H. A. &Seligman, A. M. (1953). Determinants for the specificity of action of pancreatic lipase.Arch. Biochem. Biophys. 42, 337–54.
Reid, E. & Williamson, R. (1974). Centrifugation. In:Methods in Enzymology, Vol. XXXI A (eds S. Fleischer & L. Packer), p. 714. Academic Press.
Robinson, H. (1972). An electrophoretic and biochemical analysis of acid phosphatase in the tail ofXenopus laevis during development and metamorphosis.J. exp. Zool. 180, 127–39.
Rona, P. &Takata, M. (1922). Uber die Wirkung des Chinins und der Chininderivate auf Magen lipase.Biochem. Z. 134, 118–30.
Roodyn, D. B. (1967). In:Enzyme Cytology, p. 515. Academic Press.
Stoffel, W. &Greten, H. (1967). Studies on lipolytic activities of rat liver lysosomes.Hoppe-Seyler's z. physiol. Chem. 348, 1145–50.
Underhay, E., Holt, S. J., Beaufay, H. &De Duve, C. (1956). Intracellular localisation of esterase in rat liver.J. biophys. biochem. Cytol. 2, 635–7.
Wrigglesworth, J. M. &Pover, W. F. R. (1966). Subcellular distribution of lysosomal enzymes in guinea-pig small intestine.Life Sci. 5, 1365–71.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Temple, N.J., Martin, P.A. & Connock, M.J. Zonal rotor analysis of the subcellular localization of α-glycerophosphate dehydrogenase, α-naphthyl palmitate and β-naphthyl laurate hydrolases in the mucosa of the guinea-pig small intestine. Histochem J 8, 159–175 (1976). https://doi.org/10.1007/BF01007166
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01007166