Summary
The electron microscopical localization of ornithine transcarbamylase in rat liver was investigated by a protein A—gold technique applied to thin sections of Lowicryl K4M- or LR gold-embedded materials and to ultracryosections. Gold particles were exclusively confined to mitochondria of the parenchymal cells but not of sinus-lining cells. In mitochondria, gold particles were present in the matrix and closely associated with the inner membrane. The most intensive labelling was obtained from ultracryosections, while weaker labelling was noted in sections of materials embedded in both Lowicryl K4M and LR gold. The association of the enzyme with the inner membrane was confirmed by quantitative analysis of distribution pattern.
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BENDAYAN, M., ROTH, J., PERRELET, A. & ORCI, L. (1980) Quantitative immunocytochemical localization of pancreatic secretory proteins in sub-cellular compartments of the rat acinar cell.J. Histochem. Cytochem. 28, 149–60.
BENDAYAN, M. & SHORE, G. C. (1982) Immunocytochemical localization of mitochondrial proteins in the rat hepatocyte.J. Histochem. Cytochem. 30, 139–47.
CARAVACA, J. & GRISOLIA, S. (1960) Synthesis of citrulline with animal and bacterial enzymes.J. Biol. Chem. 235, 684–93.
CLARKE, S. (1976) The polypeptides of rat liver mitochondria: identification of a 36 000 Da polypeptide as the subunit of ornithine transcarbamylase.Biochem. Biophys. Res. Commun. 71, 1118–24.
GAMBLE, J. G. & LEHNINGER, A. L. (1973) Transport of ornithine and citrulline across the mitochondrial membrane.J. Biol. Chem. 248, 610–18.
GRISOLIA, S., KORITZ, S. B. & COHEN, P. P. (1951) Dissociation of respiration and enzymatic synthesis of citrulline.J. Biol. Chem. 191, 181–7.
GRISOLIA, S. & COHEN, P. P. (1952) The catalytic role of carbamyl glutamate in citrulline biosynthesis.J. Biol. Chem. 198, 561–71.
HALL, L. M., JOHNSON, R. C. & COHEN, P. P. (1960) The presence of carbamylphosphate synthetase in intestinal mucosa.Biochim. Biophys. Acta 37, 144–5.
HOOGENRAAD, N. J., SUTHERLAND, T. M. & HOWLETT, G. J. (1980) Purification of ornithine transcarbamylase from rat liver by affinity chromatography with immobilized transition-state analog.Analyt. Biochem. 101, 97–102.
JONES, M. E., ANDERSON, A. D., ANDERSON, C. & HODES, S. (1961) Citrulline synthesis in rat tissues.Arch. Biochem. Biophys. 95, 499–507.
KALOUSEK, F., FRANCOIS, B. & ROSENBERG, L. E. (1978) Isolation and characterization of ornithine transcarbamylase from normal human liver.J. Biol. Chem. 253, 3939–44.
LAEMMLI, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–5.
LOWENSTEIN, J. M. & COHEN, P. P. (1956) Studies on the biosynthesis of carbamylaspartic acid.J. Biol. Chem. 220, 57–70.
LUSTY, C. J., JILKA, R. L. & NIETSCH, E. H. (1979) Ornithine transcarbamylase of rat liver. Kinetic, physical, and chemical properties.J. Biol. Chem. 254, 10030–6.
MARSHALL, M. & COHEN, P. P. (1972) Ornithine transcarbamylase fromStreptococcus faecalis and bovine liver.J. Biol. Chem. 247, 1641–53.
MORI, M., MIURA, S., TATIBANA, M. & COHEN, P. P. (1980) Processing of a putative precursor of rat liver ornithine transcarbamylase, a mitochondrial matrix enzyme.J. Biochem. (Tokyo) 88, 1829–36.
PIERSON, D. L., COX, S. L. & GILBERT, B. E. (1977) Human ornithine transcarbamylase. Purification and characterization of the enzyme from normal liver and the liver of a Reye's syndrome patient.J. Biol. Chem. 252, 6464–9.
RAIJMAN, L. (1974) Citrulline synthesis in rat tissues and liver content of carbamoyl phosphate and ornithine.Biochem. J. 138, 225–32.
ROTH, J., BENDAYAN, M., CARLEMALM, E., VILLIGER, W. & GARAVITO, M. (1981) The enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue.J. Histochem. Cytochem. 29, 663–71.
ROTH, J. (1982) The protein A—gold (pAg) technique. A quantitative and qualitative approach for antigen localization on thin sections. InTechniques in Immunocytochemistry (edited by BULLOCK, G. R. and PETRUSZ, P.), pp. 107–33. New York: Academic Press.
RYALL, J., NGUYEN, M., BENDAYAN, M. & SHORE, G. (1985) Expression of nuclear genes encoding the urea cycle enzymes, carbamoyl—phosphate synthase I and ornithine carbamoyl transferase, in rat liver and intestinal mucosa.Eur. J. Biochem. 152, 287–92.
SCHNAITMAN, C. & GREENAWALT, J. W. (1968) Enzymatic properties of the inner and outer membranes of rat liver mitochondria.J. Cell Biol. 38, 158–75.
SHIDA, H., GORBSKY, G., SHIDA, M. & STEINBERG, M. S. (1982) Ultrastructural and biochemical identification of ConA receptors in the desmosomes.J. Cell. Biochem. 20, 113–26.
SHIDA, H., GUIDICE, G. J., SHIDA, M., COHEN, S. M. & STEINBERG, M. S. (1984) Molecular organization of the transmembrane molecules and cytoplasmic components of the desmosome. InInternational Cell Biology 1984 (edited by SENO, S. and OKADA, Y.), p. 314, Tokyo: Japan Society for Cell Biology.
SLOT, J. W. & GEUZE, H. J. (1981) Sizing of protein A—colloidal gold probe for immunoelectron microscopy.J. Cell Biol. 90, 533–6.
STATHIS, E. C. & FABRIKANOS, S. (1958) Preparation of colloidal gold.Chem. Ind. (Lond) 27, 860–1.
TOKUYASU, K. T. (1973) A technique for ultracryotomy of cell suspensions and tissues.J. Cell Biol. 57, 551–65.
TOKUYASU, K. T. & SINGER, S. J. (1976) Improved procedures for immunoferritin labeling of ultrathin frozen sections.J. Cell Biol. 71, 894–906.
TOKUYASU, K. T. (1978) A study of positive staining of ultrathin frozen sections.J. Ultrastruct. Res. 63, 287–307.
TOWBIN, H. STAEHELIN, T. & GORDON, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proc. Natl. Acad. Sci. USA 76, 4350–4.
UCHIYAMA, C., MORI, M. & TATIBANA, M. (1981) Subcellular localization and properties onN-acetylglutamate synthesis in rat small intestinal mucosa.J. Biochem. (Tokyo) 89, 1777–86.
WEIBEL, E. D., KISTLER, G. S. & SCHERLE, W. F. (1966) Practical stereological methods for morphometric cytology.J. Cell. Biol. 30, 23–38.
YOKOTA, S. & HASHIMOTO, T. (1985) Intramitochondrial distribution of three acyl—CoA dehydrogenase in rat liver parenchymal cells revealed by quantitative immunoelectron microscopy.Acta Histochem. Cytochem. 18, 49–57.
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Yokota, S., Mori, M. Immunoelectron microscopical localization of ornithine transcarbamylase in hepatic parenchymal cells of the rat. Histochem J 18, 451–457 (1986). https://doi.org/10.1007/BF01675338
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DOI: https://doi.org/10.1007/BF01675338