Channel-Forming Abilities of Spontaneously Occurring α-Toxin Fragments from Staphylococcus aureus

• Published in: The Journal of membrane biology Vol. 234; no. 3; pp. 171 - 181
• Main Authors: Vécsey-Semjén, Beatrix, Kwak, Young-Keun, Högbom, Martin, Möllby, Roland
• Format: Journal Article
• Language: English
• Published: New York New York : Springer-Verlag 01.04.2010; Springer-Verlag
• Subjects: alpha-Toxin fragments; Animals; Bacterial Toxins - chemistry; Bacterial Toxins - metabolism; Bacterial Toxins - pharmacology; Biochemistry; Biologi; Biology; Biomedical and Life Sciences; Cell and molecular biology; Cell- och molekylärbiologi; Channel formation; Erythrocyte Membrane - metabolism; Hemolysis; Hemolysis - drug effects; Human Physiology; Humans; Hydrogen-Ion Concentration; Large unilamellar vesicles; Life Sciences; Liposomes - chemistry; Medicin och hälsovetenskap; NATURAL SCIENCES; NATURVETENSKAP; Oligomerization; Phosphatidylglycerols - chemistry; Rabbits; Staphylococcus aureus - metabolism; Unilamellar Liposomes - chemistry; Hemolysis; Channel formation; Oligomerization; Large unilamellar vesicles; α-Toxin fragments
• ISSN: 0022-2631; 1432-1424; 1432-1424
• DOI: 10.1007/s00232-010-9244-7

Pore formation by four spontaneously occurring α-toxin fragments from Staphylococcus aureus were investigated on liposome and erythrocyte membranes. All the isolated fragments bound to the different types of membranes and formed transmembrane channels in egg-phosphatidyl glycerol vesicles. Fragments of amino acids (aa) 9-293 (32 kD) and aa 13-293 (31 kD) formed heptamers, similar to the intact toxin, while the aa 72-293 (26 kD) fragment formed heptamers, octamers, and nonamers, as judged by gel electrophoresis of the liposomes. All isolated fragments induced release of chloride ions from large unilamellar vesicles. Channel formation was promoted by acidic pH and negatively charged lipid head groups. Also, the fragments' hemolytic activity was strongly decreased under neutral conditions but could be partially restored by acidification of the medium. We paid special attention to the 26-kD fragment, which, despite the loss of about one-fourth of the N-terminal part of α-toxin, did form transmembrane channels in liposomes. In light of the available data on channel formation by α-toxin, our results suggest that proteolytic degradation might be better tolerated than previously reported. Channel opening could be inhibited and open channels could be closed by zinc in the medium. Channel closure could be reversed by addition of EDTA. In contrast, digestion at the C terminus led to premature oligomerization and resulted in species with strongly diminished activity and dependent on protonation.