PMID: 3773790Jan 1, 1986Paper

Purification and characterization of heat-labile toxin from Bordetella bronchiseptica

Microbiology and Immunology
M EndohY Nakase

Abstract

The heat-labile toxin (HLT) of Bordetella bronchiseptica was purified successively from sonic extracts of phase I organisms grown in Stainer-Scholte medium, by partition in hydrophobic interaction, sucrose density gradient centrifugation, gel filtration through Sepharose 4B and 6B, isoelectric precipitation and isoelectric focusing. The purified HLT was homogeneous by disc polyacrylamide gel electrophoresis and the gel diffusion-test, and free of detectable hemagglutinin and endotoxin activity. A 386-fold purification over the crude extract was obtained at a yield of about 28%, and a minimum dose of 0.9 ng was dermonecrotizing with a lesion 5 mm in diameter in guinea pigs and induced splenoatrophy. The mouse LD50 was 200 ng (intraperitoneal) or 70 ng (intravenous). The HLT was found to be a simple protein with an isoelectric point of pI 6.9. It has a molecular weight of 102,000 estimated by Sepharose 6B gel filtration and was found to consist of two different types of polypeptide by SDS-polyacrylamide gel electrophoresis, their molecular weights being 30,000 and 20,000. Amino acid analysis showed 15 common amino acid residues, and methionine, cysteine and tryptophan were undetectable. The HLT crystallized by methylpentanediol s...Continue Reading

References

Feb 1, 1971·Journal of Medical Microbiology·T Iida, T Okonogi
Oct 1, 1970·Journal of General Microbiology·D W Stainer, M J Scholte
Sep 1, 1966·Journal of Chromatography·D JonesA N Howard
Jan 1, 1982·Pharmacology & Therapeutics·A C Wardlaw, R Parton
Feb 1, 1984·Journal of Medical Microbiology·I Livey, A C Wardlaw
Jan 1, 1984·Methods in Enzymology·G L Gilliland, D R Davies

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Citations

Jan 1, 1996·Veterinary Microbiology·A J CouttsR M Gaskell
Oct 1, 1990·Biologicals : Journal of the International Association of Biological Standardization·M EndohY Nakase
Jan 1, 1986·Microbiology and Immunology·M EndohY Nakase
Feb 15, 1994·FEMS Microbiology Letters·M NagaiY Nakase
Dec 1, 1992·Journal of Bacteriology·S Stibitz, T L Garletts
Oct 1, 1991·Infection and Immunity·Y L Zhang, R D Sekura
Sep 1, 1994·Infection and Immunity·K E Walker, A A Weiss
Dec 1, 1990·Infection and Immunity·M EndohY Nakase
Jul 1, 1991·Clinical Microbiology Reviews·B F Woolfrey, J A Moody

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