Abstract
The glycoproteins of microsomes and cytosol were studied. Various washing procedures did not release the proteins from the microsomes, and immunological tests demonstrated that the sialoproteins are not serum components. Low concentrations of deoxycholate and incubation in 0.25 M sucrose solution liberated a small amount of microsomal sialoprotein and this fraction exhibited a high degree of labeling of protein-bound N-acetylneuraminic acid. A part of the glycoprotein fraction could not be solubilized, even with a high concentration of the detergent. Thoroughly perfused rat liver contained sialoproteins in the particle-free supernate. The level of sialoprotein present could not be due to contamination with serum or broken organelles. The high in vivo incorporation of [3H]glucosamine into protein-bound sialic acid of Golgi membranes and cytosol was paralleled by a delayed and lesser rate of incorporation into the rough and smooth microsomal membranes. This incorporation pattern suggests the possibility that the glycoproteins of cytosol and Golgi may later be incorporated into the membrane of the endoplasmic reticulum.
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- Autuori F., Svensson H., Dallner G. Biogenesis of microsomal membrane glycoproteins in rat liver. II. Purification of soluble glycoproteins and their incorporation into microsomal membranes. J Cell Biol. 1975 Dec;67(3):700–714. doi: 10.1083/jcb.67.3.700. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Autuori F., Svensson H., Dallner G. Relationship of protein-bound sialic acid to microsomal membranes. Biochem Biophys Res Commun. 1974 Feb 27;56(4):1023–1030. doi: 10.1016/s0006-291x(74)80291-3. [DOI] [PubMed] [Google Scholar]
- Avrameas S., Ternynck T. The cross-linking of proteins with glutaraldehyde and its use for the preparation of immunoadsorbents. Immunochemistry. 1969 Jan;6(1):53–66. doi: 10.1016/0019-2791(69)90178-5. [DOI] [PubMed] [Google Scholar]
- Beesley R. C., Forte J. G. Glycoproteins and glycolipids of oxyntic cell microsomes. I. Glycoproteins: carbohydrate composition, analytical and preparative fractionation. Biochim Biophys Acta. 1973 May 11;307(2):372–385. doi: 10.1016/0005-2736(73)90103-x. [DOI] [PubMed] [Google Scholar]
- Bizzi A., Marsh J. B. Further observations on the attachment of carbohydrate to lipoproteins by rat liver Golgi membranes. Proc Soc Exp Biol Med. 1973 Dec;144(3):762–765. doi: 10.3181/00379727-144-37677. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B., Hagopian A., Eylar E. H. Cellular membranes: the biosynthesis of glycoprotein and glycolipid in hela cell membranes. Arch Biochem Biophys. 1969 Mar;130(1):573–583. doi: 10.1016/0003-9861(69)90073-3. [DOI] [PubMed] [Google Scholar]
- Bosmann H. B. Synthesis of glycoproteins in brain: identification, purification and properties of a synaptosomal sialyl transferase utilizing endogenous and exogenous acceptors. J Neurochem. 1973 Apr;20(4):1037–1049. doi: 10.1111/j.1471-4159.1973.tb00075.x. [DOI] [PubMed] [Google Scholar]
- Dallner G., Azzi A. Structural properties of rough and smooth microsomal membranes. A study with fluorescence probes. Biochim Biophys Acta. 1972 Feb 11;255(2):589–601. doi: 10.1016/0005-2736(72)90163-0. [DOI] [PubMed] [Google Scholar]
- Dallner G. Isolation of rough and smooth microsomes--general. Methods Enzymol. 1974;31:191–201. [PubMed] [Google Scholar]
- Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. I. Structural and chemical differentiation in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):73–96. doi: 10.1083/jcb.30.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorling P. R., Le Page R. N. A rapid high yield method for the preparation of rat liver cell plasma membranes. Biochim Biophys Acta. 1973 Aug 9;318(1):33–40. doi: 10.1016/0005-2736(73)90333-7. [DOI] [PubMed] [Google Scholar]
- ERNSTER L., LOW H. Reconstruction of oxidative phosphorylation in aged mitochondrial systems. Exp Cell Res. 1955;(Suppl 3):133–153. [PubMed] [Google Scholar]
- Ehrenreich J. H., Bergeron J. J., Siekevitz P., Palade G. E. Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization. J Cell Biol. 1973 Oct;59(1):45–72. doi: 10.1083/jcb.59.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Evans W. H., Gurd J. W. Biosynthesis of liver membranes. Incorporation of ( 3 H)leucine into proteins and of ( 14 C)glucosamine into proteins and lipids of liver microsomal and plasma-membrane fractions. Biochem J. 1971 Nov;125(2):615–624. doi: 10.1042/bj1250615. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eylar E. H. On the biological role of glycoproteins. J Theor Biol. 1966 Jan;10(1):89–113. doi: 10.1016/0022-5193(66)90179-2. [DOI] [PubMed] [Google Scholar]
- Glaumann H., Dallner G. Lipid composition and turnover of rough and smooth microsomal membranes in rat liver. J Lipid Res. 1968 Nov;9(6):720–729. [PubMed] [Google Scholar]
- Glick M. C., Comstock C. A., Cohen M. A., Warren L. Membranes of animal cells. 8. Distribution of sialic acid, hexosamines and sialidase in the L cell. Biochim Biophys Acta. 1971 Apr 13;233(2):247–257. doi: 10.1016/0005-2736(71)90324-5. [DOI] [PubMed] [Google Scholar]
- Goldstone A., Koenig H., Nayyar R., Hughes C., Lu C. Y. Isolation and characterization of a rough microsomal fraction from rat kidney that is enriched in lysosomal enzymes. Biochem J. 1973 Feb;132(2):259–266. doi: 10.1042/bj1320259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldstone A., Koenig H. Synthesis and turnover of lysosomal glycoproteins. Relation to the molecular heterogeneity of the lysosomal enzymes. FEBS Lett. 1974 Feb 15;39(2):176–181. doi: 10.1016/0014-5793(74)80045-1. [DOI] [PubMed] [Google Scholar]
- Harms E., Reutter W. Half-life of N-acetylneuraminic acid in plasma membranes of rat liver and Morris hepatoma 7777. Cancer Res. 1974 Dec;34(12):3165–3172. [PubMed] [Google Scholar]
- Helgeland L., Christensen T. B., Janson T. L. The distribution of protein-bound carbohydrates in submicrosomal fractions from rat liver. Biochim Biophys Acta. 1972 Nov 24;286(1):62–71. doi: 10.1016/0304-4165(72)90088-8. [DOI] [PubMed] [Google Scholar]
- Kawasaki T., Yamashina I. Isolation and characterization of glycopeptides from rat liver nuclear membrane. J Biochem. 1972 Dec;72(6):1517–1525. doi: 10.1093/oxfordjournals.jbchem.a130043. [DOI] [PubMed] [Google Scholar]
- Kawasaki T., Yamashina I. Metabolic studies of rat liver plasma membranes using D-(1-14C)glucosamine. Biochim Biophys Acta. 1971 Feb 2;225(2):234–238. doi: 10.1016/0005-2736(71)90216-1. [DOI] [PubMed] [Google Scholar]
- Kim Y. S., Perdomo J. M. Membrane glycoproteins of the rat small intestine. Chemical composition of membrane glycoproteins. Biochim Biophys Acta. 1974 Mar 14;342(1):111–124. doi: 10.1016/0005-2795(74)90112-3. [DOI] [PubMed] [Google Scholar]
- Kreibich G., Sabatini D. D. Selective release of content from microsomal vesicles without membrane disassembly. II. Electrophoretic and immunological characterization of microsomal subfractions. J Cell Biol. 1974 Jun;61(3):789–807. doi: 10.1083/jcb.61.3.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Leighton F., Poole B., Beaufay H., Baudhuin P., Coffey J. W., Fowler S., De Duve C. The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions. J Cell Biol. 1968 May;37(2):482–513. doi: 10.1083/jcb.37.2.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martin S. S., Bosmann H. B. Glycoprotein nature of mitochondrial structural protein and neutral sugar content of mitochondrial proteins and structural proteins. Exp Cell Res. 1971 May;66(1):59–64. doi: 10.1016/s0014-4827(71)80010-1. [DOI] [PubMed] [Google Scholar]
- Miyajima N., Tomikawa M., Kawasaki T., Yamashina I. Chemical composition of membranous fractions of rat liver microsomes. J Biochem. 1969 Nov;66(5):711–732. [PubMed] [Google Scholar]
- Molnar J. Glycoproteins of Ehrlich ascites carcinoma cells. Incorporation of [14C]glucosamine and [14C]sialic acid into membrane proteins. Biochemistry. 1967 Oct;6(10):3064–3076. doi: 10.1021/bi00862a013. [DOI] [PubMed] [Google Scholar]
- Pepper D. S., Jamieson G. A. Studies on glycoproteins. 3. Isolation of sialylglycopeptides from human platelet membranes. Biochemistry. 1969 Aug;8(8):3362–3369. doi: 10.1021/bi00836a034. [DOI] [PubMed] [Google Scholar]
- Quirk S. J., Byrne J., Robinson G. B. Studies on the turnover of protein and glycoprotein components in rabbit kidney brush borders. Biochem J. 1973 Mar;132(3):501–508. doi: 10.1042/bj1320501. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Redman C. M., Cherian M. G. The secretory pathways of rat serum glycoproteins and albumin. Localization of newly formed proteins within the endoplasmic reticulum. J Cell Biol. 1972 Feb;52(2):231–245. doi: 10.1083/jcb.52.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schachter H., Jabbal I., Hudgin R. L., Pinteric L., McGuire E. J., Roseman S. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction. J Biol Chem. 1970 Mar 10;245(5):1090–1100. [PubMed] [Google Scholar]
- Simkin J. L., Jamieson J. C. Studies on the site of biosynthesis of acidic glycoproteins of guinea-pig serum. Biochem J. 1967 Apr;103(1):153–164. doi: 10.1042/bj1030153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]