Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Immunochemical analysis of the carbohydrate moiety of yeast killer toxin K28

  • 45 Accesses

  • 5 Citations


Killer toxin K28, a 16 kd protein secreted by the wine yeast Saccharomyces cerevisiae strain 28, was reversibly bound by a column of Concanavalin A-Sepharose, confirming its glycoprotein nature. HPLC analysis of acid hydrolyzates of K28 toxin as well as Western-blots of β-eliminated and/or endo H-treated killer toxin preparations probed with polyclonal α-toxin antibodies revealed that the carbohydrate moiety of K28 consists of D-mannose only, which is O-glycosidically linked via Ser/Thr residues to the protein part. The change in gel mobility of K28 after β-elimination was caused by a decrease in molecular mass of about 1,800, corresponding to a carbohydrate moiety of 10 mannose residues per killer toxin molecule.

This is a preview of subscription content, log in to check access.


  1. Bostian KA, Elliott Q, Bussey H, Burn V, Smith A & Tipper DJ (1984) Sequence of the preprotoxin dsRNA gene of type 1 killer yeast: Multiple processing events produce a two-component toxin. Cell 36: 741–751

  2. De la Pena P, Barros F, Gascon S, Lazo PS & Ramos S (1981) Effect of yeast killer toxin on sensitive cells of Saccharomyces cerevisiae. J. Biol. Chem. 256: 10420–10425

  3. Dresser DW (1986) Immunization of experimental animals. In: Weir M (ed) Handbook of Experimental Immunology Vol 1, Immunochemistry. Blackwell Scientific Publications, Oxford

  4. Hutchins K & Bussey H (1983) Cell wall receptor for yeast killer toxin: involvement of β-1, 6-D-glucan. J. Bacteriol. 154: 161–169

  5. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

  6. Manners DJ, Masson AJ & Patterson JC (1973) The structure of a β-1, 3-D-glucan from yeast cell walls. Biochem. J. 135: 19–30

  7. Montreuil J, Bouquelet S, Debray H, Fournet B, Spik G & Strecker G (1986), Glycoproteins. In: Chaplin MF & Kennedy JF (Eds) Carbohydrate Analysis. A Practical Approach. IRL Press Oxford

  8. Nakajima T & Ballou CE (1974) Characterization of the carbohydrate fragments obtained from Saccharomyces cerevisiae mannan by alkaline degradation. J. Biol. Chem. 249: 7679–7684

  9. Palacios R, Palmiter RD & Schimke RT (1972) Identification and isolation of ovalbumin-synthesizing polysomes. 1. Specific binding of 125I-anti-ovalbumin to polysomes. J. Biol. Chem. 247: 2316–2321

  10. Pfeiffer P & Radler F (1982) Purification and characterization of extracellular and intracellular killer toxin of Saccharomyces cerevisiae strain 28. J. Gen. Microbiol. 128: 2699–2706

  11. Pfeiffer P & Radler F (1984) Comparison of the killer toxin of several yeasts and purification of a toxin of type K2. Arch. Microbiol. 137: 357–361

  12. Pfeiffer P & Radler F (1985) Hochleistungsflüssigchromatographische Bestimmung von organischen Säuren, Zuckern, Glycerin und Alkohol im Wein an einer Kationenaustauschersäule. Z. Lebens. Unters. Forsch. 181: 24–27

  13. Schmitt M, Brendel M, Schwarz R & Radler F (1989) Inhibition of DNA synthesis in Saccharomyces cerevisiae by yeast killer toxin KT28. J. Gen. Microbiol. 135: 1529–1535

  14. Schmitt M & Radler F (1987) Mannoprotein of the yeast cell wall as primary receptor for the killer toxin of Saccharomyces cerevisiae strain 28. J. Gen. Microbiol. 133: 3347–3354

  15. Schmitt M & Radler F (1989) Purification of yeast killer toxin KT28 by receptor-mediated affinity chromatography. J. Chromatogr. 469: 448–452

  16. Schmitt MJ & Tipper DJ (1990) K28: A unique double-stranded RNA killer virus of Saccharomyces cerevisiae. Mol. Cell. Biol. 10: 4807–4815

  17. Tipper DJ & Bostian KA (1984) Double-stranded ribonucleic acid killer systems in yeasts. Microbiol. Rev. 48: 125–156

  18. 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: 4350–4354

  19. Wickner RB (1986) Double-stranded RNA replication in yeast: the killer system. Ann. Rev. Biochem. 55: 373–395

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Schmitt, M.J., Pfeiffer, P.C. Immunochemical analysis of the carbohydrate moiety of yeast killer toxin K28 . Antonie van Leeuwenhoek 58, 277–282 (1990).

Download citation

Key words

  • Saccharomyces cerevisiae
  • killer toxin K28
  • glycoprotein
  • O-glycosylation
  • β-elimination
  • α-toxin antibodies