Purification of alpha-toxin from Staphylococcus aureus and application to cell permeabilization - PubMed (original) (raw)
Purification of alpha-toxin from Staphylococcus aureus and application to cell permeabilization
I Lind et al. Anal Biochem. 1987 Jul.
Abstract
Crude alpha-toxin was produced by Staphylococcus aureus, strain Wood 46. The amount of exotoxin was monitored during growth and all subsequent purification steps by determination of its hemolytic activity against rabbit erythrocytes. The culture supernatant was treated with ammonium sulfate (75% saturation). The resulting precipitate was dialyzed and subjected to cation-exchange chromatography. The fractions containing the hemolytic activity were further purified by gel chromatography. The final product was enriched by a factor of 8.5 compared to the crude toxin. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified toxin exhibited one major band. It caused the release of 86Rb+ and ATP from rat insulinoma (RIN A2) as well as pheochromocytoma cells (PC12) in culture, indicating efficient permeabilization of their plasma membranes for small molecules.
Similar articles
- Minimal requirements for exocytosis. A study using PC 12 cells permeabilized with staphylococcal alpha-toxin.
Ahnert-Hilger G, Bhakdi S, Gratzl M. Ahnert-Hilger G, et al. J Biol Chem. 1985 Oct 15;260(23):12730-4. J Biol Chem. 1985. PMID: 4044606 - Purification of oligomeric staphylococcal alpha-toxin by affinity chromatography on digitonin-sepharose.
Schaeg W, Kranz T, Brückler J, Blobel H. Schaeg W, et al. Zentralbl Bakteriol Mikrobiol Hyg A. 1985 Feb;259(1):71-7. doi: 10.1016/s0176-6724(85)80009-2. Zentralbl Bakteriol Mikrobiol Hyg A. 1985. PMID: 4002934 - Purification and characterization of staphylococcal pyrogenic exotoxin type B.
Schlievert PM. Schlievert PM. Biochemistry. 1980 Dec 23;19(26):6204-8. doi: 10.1021/bi00567a040. Biochemistry. 1980. PMID: 6781533 - Non-thiol-activated cytolytic bacterial toxins: current status.
Kato I, Morinaga N, Muneto R. Kato I, et al. Microbiol Sci. 1988 Feb;5(2):53-7. Microbiol Sci. 1988. PMID: 3152763 Review. - [Molecular cellular aspects of the hemolytic action of staphylococcal alpha-toxin].
Ivanov NR, Brill' GE. Ivanov NR, et al. Zh Mikrobiol Epidemiol Immunobiol. 1984 Sep;(9):3-9. Zh Mikrobiol Epidemiol Immunobiol. 1984. PMID: 6391044 Review. Russian. No abstract available.
Cited by
- Functional localization of an exocytosis-triggering G-protein in human cytotoxic T lymphocytes.
Mittrücker HW, Fleischer B. Mittrücker HW, et al. Immunology. 1992 Aug;76(4):610-5. Immunology. 1992. PMID: 1383135 Free PMC article. - Changes in the mechanism of Ca2(+) mobilization during the differentiation of BC3H1 muscle cells.
De Smedt H, Parys JB, Himpens B, Missiaen L, Borghgraef R. De Smedt H, et al. Biochem J. 1991 Jan 1;273(Pt 1)(Pt 1):219-23. doi: 10.1042/bj2730219. Biochem J. 1991. PMID: 1989585 Free PMC article. - Rho-kinase inhibition attenuates calcium-induced contraction in β-escin but not Triton X-100 permeabilized rabbit femoral artery.
Clelland LJ, Browne BM, Alvarez SM, Miner AS, Ratz PH. Clelland LJ, et al. J Muscle Res Cell Motil. 2011 Sep;32(2):77-88. doi: 10.1007/s10974-011-9253-x. Epub 2011 Jun 25. J Muscle Res Cell Motil. 2011. PMID: 21706258 Free PMC article. - A Wall Fragment of Cutibacterium acnes Preserves Junctional Integrity Altered by Staphylococcus aureus in an Ex Vivo Porcine Skin Model.
Magnifico I, Perna A, Cutuli MA, Medoro A, Pietrangelo L, Guarnieri A, Foderà E, Passarella D, Venditti N, Vergalito F, Petronio Petronio G, Di Marco R. Magnifico I, et al. Pharmaceutics. 2023 Apr 12;15(4):1224. doi: 10.3390/pharmaceutics15041224. Pharmaceutics. 2023. PMID: 37111709 Free PMC article. - Alpha-toxin of Staphylococcus aureus.
Bhakdi S, Tranum-Jensen J. Bhakdi S, et al. Microbiol Rev. 1991 Dec;55(4):733-51. doi: 10.1128/mr.55.4.733-751.1991. Microbiol Rev. 1991. PMID: 1779933 Free PMC article. Review.