Skip to main content
Log in

Immunochemical analysis of the carbohydrate moiety of yeast killer toxin K28

  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

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 via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • 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

    Article  PubMed  Google Scholar 

  • 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

    PubMed  Google Scholar 

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

    Google Scholar 

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

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • 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

    PubMed  Google Scholar 

  • 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

    Google Scholar 

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

    PubMed  Google Scholar 

  • 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

    PubMed  Google Scholar 

  • 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

    Google Scholar 

  • 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

    PubMed  Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    PubMed  Google Scholar 

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

    Article  Google Scholar 

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

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • 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

    PubMed  Google Scholar 

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

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

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). https://doi.org/10.1007/BF00399340

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00399340

Key words

Navigation