The presence of glycopeptide lectin receptors in the ascitic fluid of rats bearing Novikoff or AS-30D hepatoma was investigated. Macrosialoglycopeptides, resistant to pronase digestion, were partially purified from the ascitic fluid of hepatoma-bearing rats by gel filtration on Sephadex G-50. A macrosialoglycopeptide fraction, isolated from the ascitic fluid of rats bearing the Novikoff hepatoma, possessed potent concanavalin A (Con A) receptor activity. This fraction possessed higher Con A receptor activity than did the comparable macrosialoglycopeptide fraction from the ascitic fluid of rats bearing the AS-30D hepatoma; this observation is in agreement with the Con A-induced agglutination properties of these 2 hepatoma cell lines and with the Con A receptor activities of the glycopeptides released from the surface of the hepatoma cells by papain digestion. Rat blood serum contained a comparable macrosialoglycopeptide fraction, which possessed weak Con A receptor activity. The macrosialoglycopeptide fractions from the ascitic fluid of hepatoma-bearing rats possessed wheat germ agglutinin receptor activity. However, this activity was also present in normal rat serum. These results suggest that glycopeptides present on the surface of Novikoff hepatoma cells are shed into the ascitic fluid and may be distinguished from components in normal serum by their Con A receptor activity.
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References
1.
AdairW.L., KornfeldS.: Isolation of the receptors for wheat germ agglutinin and the Ricinus Communis lectins from human erythrocytes using affinity chromatography.J. Biol. Chem.,249: 4696–4704, 1974.
2.
AminoffD.: Methods for the quantitative estimation of N-acetyl-neuraminic acid and their application to hydrolyzates of sialomucins.Biochem. J.,81: 384–392, 1961.
3.
BhattiT., ClampJ.R.: Determination of molecular weight of glycopeptides by exclusion chromatography.Biochim. Biophys. Acta,170: 206–208, 1968.
CooperA.G., CodingtonJ.F., BrownM.C.: In vivo release of glycoprotein I from the Ha subline of TA3 murine tumor into ascites fluid and serum.Proc. Natl. Acad. Sci. U.S.A.,71: 1224–1228, 1974.
6.
Di FerranteN.: Measurement of urinary mucopolysaccharides.Anal. Biochem.,21: 98–106, 1967.
7.
DoljanskiF.A.: A new look at the cell surface.Israel J. Med. Sci.,9: 251–257, 1973.
8.
DuboisM., GillesK.A., HamiltonJ.K., RebersP.A., SmithF.: Colorimetric method for the determination of sugars and related substances.Anal. Chem.,28: 350–354, 1956.
9.
EngenR.L.: Effect of steroid on serum sialic acid values in the canine.Proc. Soc. Exp. Biol. Med.,135: 778–780, 1970.
10.
GattR., BermanE.R.: A rapid procedure for estimation of amino sugars on a micro scale.Anal. Biochem.,15: 167–171, 1966.
11.
HuntR.C., BullisC.M., BrownJ.C.: Isolation of a concanavalin A receptor from mouse L cells.Biochemistry,14: 109–115, 1975.
12.
JansonV.K., BurgerM.M.: Isolation and characterization of agglutinin receptor sites. II. Isolation and partial purification of a surface membrane receptor for wheat germ agglutinin.Biochim. Biophys. Acta,291: 128–135, 1973.
13.
KapellerM., Gal-OzR., GroverN.B., DoljanskiF.: Natural shedding of carbohydrate-containing macromolecules from cell surfaces.Exp. Cell Res.,79: 152–158, 1973.
14.
LeseneyA.M., ThomasM.W., O'NeillP.A., WalborgE.F.Jr.: Role of papain-labile, cell-surface glycopeptides in agglutination of rat erythrocytes by concanavalin A and wheat germ agglutinin.Colloques internationaux N° 221 du Centre National de la Recherche Scientifique sur les glycoconjuguès, Vol. 2, pp. 859–868. Editions du C.N.R.S., Paris, 1974.
15.
MontgomeryR., LeeY.C., WuY.C.: Glycopeptides from ovalbumin. Preparation, properties and partial hydrolysis of the asparaginyl carbohydrate.Biochemistry,4: 566–577, 1965.
NeriG., SmithD.F., GilliamE.B., WalborgE.F.Jr.: Concanavalin A and wheat germ agglutinin receptor activity of sialoglycopeptides isolated from the surface of Novikoff hepatoma cells.Arch. Biochem. Biophys.,165: 323–330, 1974.
18.
NeriG., SmithD.F., GilliamE.B., WalborgE.F.Jr.: Concanavalin A and wheat germ agglutinin receptor activity of sialoglycopeptides isolated from the surface of rat hepatoma cells. pp. 41–48. In: Comparative Biochemistry and Physiology of Transport.BolisL., BlockK., LuriaS.E., LynenF. (eds.). North Holland Publishing Co., Amsterdam, 1974.
19.
NicolsonG.L.: The interactions of lectins with animal cell surfaces.Int. Rev. Cytol.,39: 89–190, 1974.
20.
NovikoffA.B.: A transplantable rat liver tumor induced by 4-dimethyl-aminoazo-benzene.Cancer Res.,17: 1010–1027, 1957.
21.
PepperD.S., JamiesonG.A.: Isolation of a macrosialoglycopeptide from human platelets.Biochemistry,9: 3706–3713, 1970.
22.
SmithD.F., NeriG., WalborgE.F.Jr.: Isolation and partial characterization of cell-surface glycopeptides from AS-30D rat hepatoma which possess binding sites for wheat germ agglutinin and concanavalin A.Biochemistry,12: 2111–2118, 1973.
23.
SmithD.F., WalborgE.F.Jr.: Isolation and chemical characterization of cell-surface sialoglycopeptide fractions during progression of rat ascites hepatoma AS-30D.Cancer Res.,32: 543–545, 1972.
24.
SmithD.F., WalborgE.F.Jr., ChangJ.P.: Establishment of a transplantable ascites variant of a rat hepatoma induced by 3'-methyl-4-dimethyl-aminoazobenzene.Cancer Res.,30: 2306–2309, 1970.
25.
StarlingJ.J., CapetilloS.C., NeriG., WalborgE.F.Jr.: Surface properties of normal and neoplastic rat liver cells. Lectin-induced cytoagglutination and lectin receptor activity of cell-surface glycopeptides.Exp. Cell Res.,104: 177–190, 1977.
26.
Van BlitterswijkW.J., EmmelotP., HilgerJ., KamlagD., NusseR., FeltkampC.A.: Quantitation of virus-induced (MLr) and normal (Thy. 1.2) cell surface antigens in isolated plasma membranes and the extracellular ascites fluid of mouse leukemia cells.Cancer Res.,35: 2743–2751, 1975.
27.
WalborgE.F.Jr., LantzR.S., WrayV.P.: Isolation and chemical characterization of a cell-surface sialoglycopeptide fraction from Novikoff ascites cells.Cancer Res.,29: 2034–2038, 1969.
28.
WalborgE.F., WrayV.P., SmithD.F., NeriG.: Alterations of cell-surface glycoproteins during malignant transformation: Receptor for the phytoagglutinins, concanavalin A and wheat germ agglutinin. pp. 123–147. In: Immunological Aspects of Neoplasia. University of Texas System Cancer Center M.D. Anderson Hospital and Tumor Institute. The Williams and Wilkins Co., Baltimore, 1975.
29.
WarrenL.: The biological significance of turnover of the surface membrane of animal cells. Vol. IV, pp. 197–222. In: Current Topics in Development Biology.MosconaA. A., MonroyA. (eds.). Academic Press, New York, 1969.
30.
WinzlerR.J.: Glycoproteins of plasma membranes. Chemistry and function. Part. B., pp. 1268–1293. In: Glycoproteins. Their Composition, Structure and Function.GottschalkA. (ed.). Elsevier Publishing Co., New York, 1972.
31.
WrayV.P., WalborgE.F.Jr.: Isolation of tumor cell surface binding sites for concanavalin A and wheat germ agglutinin.Cancer Res.,31: 2072–2079, 1971.
