EurekaMag
+ Translate
+ Most Popular
Spermatogenic and steroidogenic impairment after chromium treatment in rats
Prediction of enthalpy of formation based on refined crystal structures of multisite compounds; Part 1, Theories and examples
Intrastromal photorefractive keratectomy for myopia by Nd:YLF picosecond laser
Teachers' beliefs about the role of prior language knowledge in learning and how these influence teaching practices
Classification and genesis of submarine iron-manganese deposits
Insecta a Cl. G. N. Potanin in China et in Mongolia novissime lecta. XVII. Hemiptera-Heteroptera
On the occurrence of the splendid genus Machaerothrix Haupt in Tsushima, with some remarks on the genus (Hymenoptera, Pompilidae)
Enamel craze lines
Gynecologic examination of the child and the adolescent
Periosteal osteoclasts, skeletal development and ageing
Fibrodysplasia ossificans progressiva in a Nigerian: a case report
The chronically ill patient with anorexia nervosa development, phenomenology, and therapeutic considerations
Kummelonautilus taiwanum sp. nov. (Nautilida) from the early Miocene Shihmen Formation, Chungliao, Nantou, west-central Taiwan
Efficacy of Droncit against Dipylidium caninum in dogs
Pierids of the genus Euchloe Klots (Lepidoptera, Pieridae) from Siberia and the Far East.
C5A anaphylatoxin and its seven transmembrane-segment receptor
Failure to practice evidence-based medicine: why do physicians not treat patients with heart failure with angiotensin-converting enzyme inhibitors?
Empirical comparisons of proportional hazards regression, Poisson regression, and logistic regression in the analysis of occupational cohort data
Pollination of flowers of sweet and sour cherries
Isopodi terrestri raccolti nell' Estremo Oriente
Morphometric observations on Dipylidium caninum from dogs and cats in Austria
Chelation effectiveness of zinc proteinates demonstrated
Alopecia--a psychosomatic disease picture? I. Review of the literature
Monitoring equity in health and healthcare: a conceptual framework
Hemodynamic evaluation of the Biocor heterologous prosthesis

Effect of trypsin on the cell surface proteins of hepatoma tissue culture cells. Characterization of a carbohydrate-rich glycopeptide released from a calcium binding membrane glycoprotein


Effect of trypsin on the cell surface proteins of hepatoma tissue culture cells. Characterization of a carbohydrate-rich glycopeptide released from a calcium binding membrane glycoprotein



Journal of Biological Chemistry 254(10): 3935-3946



ISSN/ISBN: 0021-9258

PMID: 438168

Concentrations of trypsin that bring about aggregation of hepatoma tissue culture (HTC) cells also release from the cell surface an Mr = 55,000 glycopeptide fragment. This glycopeptide fragment also accumulates in the medium, including serum-free medium, as a normal consequence of membrane protein turnover. The trypsin-released glycopeptide is labeled when cells are grown in the presence of fucose or leucine before treatment of the cells with the protease. Similarly, the glycopeptide fragment can be labeled by reacting cells in situ by lactoperoxidase-catalyzed radioiodination or by tritiated borohydride reduction of cells treated first with neuraminidase and galactose oxidase. The tryptic glycopeptide fragment was purified by concanavalin A-Sepharose chromatography, and hydroxyapatite chromatography in the presence of dodecyl sulfate. The amino acid and carbohydrate composition was determined, as was the sensitivity of the purified glycopeptide to a variety of endo- and exoglycosidases. The purified glycopeptide contains an average of 17 sialic acid residues and hence, shows charge heterogeneity after electrophoresis in isoelectric focusing gels. The charge heterogeneity can be eliminated completely by treatment with neuraminidase. The glycopeptide after this treatment is homogeneous. The trypsin-sensitive membrane glycoprotein which is the source of the Mr = 55,000 glycopeptide was identified by two-dimensional gel electrophoretic analysis of labeled cells, treated or not treated with trypsin. This glycoprotein, which has an apparent molecular weight of 85,000 and forms a homodimer in the presence of calcium ions, was purified and its identity as the parent of the Mr = 55,000 glycopeptide was confirmed by showing that the same Mr = 55,000 fragment was released by trypsin from the purified glycoprotein as was released from the intact cells.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 039947552

Download citation: RISBibTeXText

Related references

Cell surface radiolabeling of the carbohydrate moieties of the plasma membrane major glycoprotein of AH-66 hepatoma ascites cells. Chemical and Pharmaceutical Bulletin 31(6): 2039-2044, 1983

Purification and characterization of a major glycoprotein in rat hepatoma plasma membranes. One of the membrane proteins released by phosphatidylinositol-specific phospholipase C. Biochemical Journal 241(1): 63-70, 1987

Purification and characterization of a major glycoprotein in rat hepatoma plasma membranes: one of the membrane proteins released by phosphatidylinositol-specific phospholipase C. Biochemical Journal (London 1906) 241(1): 63-70, 1987

The C1q-binding cell membrane proteins cC1q-R and gC1q-R are released from activated cells: Subcellular distribution and immunochemical characterization. Clinical Immunology and Immunopathology 84(1): 17-26, 1997

Studies on rat ascites hepatoma cells in tissue culture. I. Trials to establish a cell strain in tissue culture, especially on the growth-promoting substances for hepatoma cells in earlier stages of passage culture. Gan 48(4): 532-534, 1957

Expression and characterization of a lactosaminoglycan-carrying glycoprotein of Zajdela hepatoma cell surface--structural analysis of the carbohydrate moiety. European Journal of Biochemistry 247(3): 1091-1101, 1997

Proteins of the hepatoma tissue culture cell plasma membrane. Journal of Supramolecular Structure 4(2): 141-159, 1976

Turnover of the plasma membrane proteins of hepatoma tissue culture cells. Journal of Biological Chemistry 251(3): 872-882, 1976

Characterization of cell-surface material released from AH-66 hepatoma ascites cells into an isotonic solution. Chemical and Pharmaceutical Bulletin 34(1): 206-213, 1986

Turnover of iodinated plasma membrane proteins of hepatoma tissue culture cells. Federation Proceedings 33(5 Part 2): 1415, 1974

The Streptococcus gordonii surface proteins GspB and Hsa mediate binding to sialylated carbohydrate epitopes on the platelet membrane glycoprotein Ibalpha. Infection and Immunity 72(11): 6528-6537, 2004

Enzyme expression and growth control in hepatoma tissue culture cells and hybrids of hepatoma tissue culture cells with other cell types. Davidson, Richard L And Felix F De La Cruz (Ed ) Somatic Cell Hybridization Proceedings Of A Conference Winter Park, Fla , U S A , March 11-14, 1973 Xvii+295p Illus Raven Press: New York, N Y , U S A ISBN 0-911216-75-8: 179-184, 1974

Purification and some properties of cyclic amp binding proteins from rat liver and hepatoma tissue culture cells. Federation Proceedings 35(7): 1805, 1976

A hydroxyproline-containing glycopeptide released from the walls of sycamore tissue-culture cells by hydrazinolysis. Biochemical Journal 135(2): 327-329, 1973

Isolation and characterization of an amino sugar-rich glycopeptide from the surface layer glycoprotein of Thermoanaerobacterium thermosaccharolyticum E207-71. Carbohydrate Research 295: 245-253, 1996