Purification and some properties of a lethal toxic fragment of staphylococcal α-toxin by tryptic digestion

Watanabe, Masashi; Kato, Ikunoshin

doi:10.1016/0005-2795(78)90104-6

Cited by 21 publications

(10 citation statements)

References 19 publications

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“…(i) The a-toxin region(s) responsible for hemolysis and dermo-necrosis (present in a-13) is separate from the region responsible for lethality (lacking from a-13). This is in agreement with earlier findings (7,19). (ii) The hemolytic and dermo-necrotic region(s) is also separate from the region responsible for intoxication of Y 1 cells.…”

Section: Discussionsupporting

confidence: 93%

“…We have no amino acid sequence data for the a-1 3 fragment, but since it is non-lethal it should lack at least part of the region corresponding to the lethal fragment. The following principal conclusions can be made: (7,19). It is assumed that a-I3 resulted from cleavage of a-toxin at only one position.…”

Section: Discussionmentioning

confidence: 99%

“…intact a-toxin, the a-13 fragment and two previously described tryptic fragments (7,19). A 17 000 M, fragment (heavy chain, H.C.) was lethal "'I 28 HU/ml of a-13 were incubated at 22 "C for 30 min with polyclonal rabbit anti-a-toxin diluted as indicated.…”

Section: Lack Of Lethal Activity Of A-i3mentioning

confidence: 99%

“…We reported similar findings regarding the interaction of a-toxin with Y 1 cells (1 3). Moreover, a lethal but non-hemolytic and non-dermo-necrotic tryptic fragment (I7 000 M,) of a-toxin has been described (7,19), demonstrating separate regions for lethality and hemolysis.…”

mentioning

confidence: 99%

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A Staphylococcal Α‐toxin Fragment

Blomqvist

Thelestam

1986

Acta Pathologica Microbiologica Scandinavica Series B: Microbio

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A fragment (α‐13) of Staphylococcal α‐toxin was compared with intact α‐toxin as regards biochemical and biological properties, and the resulting information was used for mapping biologically‐active regions of α‐toxin. The α‐13 fragment had an apparent Mr of 18 500, judged by sodium dodecylsulphate polyacrylamide gel electrophoresis. However, it showed the same relative mobility as α‐toxin when subjected to gel filtration on Biogel P60, high pressure liquid chromatography or electrophoresis on polyacrylamide gradient gel. The fragment had roughly the same specific hemolytic activity as intact α‐toxin. In contrast to intact α‐toxin, the fragment was neither membrane‐damaging to mouse adrenocortical (Y 1) tumour cells nor lethal to mice. However, a short treatment of Y 1 cells with the fragment completely blocked intoxication by subsequently‐added α‐toxin. Likewise, the lethal effect of α‐toxin was inhibited when the fragment was injected prior to the toxin. Thus, the fragment had lost the active region(s) responsible for Y 1 cell intoxication and lethality, while the region(s) for binding to these targets, as well as the region responsible for hemolysis, were retained. On the basis of these findings and previous reports concerning tryptic fragments of α‐toxin, a hypothetical map of the different biologically‐active regions of α‐toxin was established.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Lack Of Lethal Activity Of A-i3mentioning

confidence: 99%

mentioning

confidence: 99%

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A Staphylococcal Α‐toxin Fragment

Blomqvist

Thelestam

1986

Acta Pathologica Microbiologica Scandinavica Series B: Microbio

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“…Oxy-haemoglobin (oxy-Hb) was prepared as described by Martin, Villani, Jothianandan & Furchgott (1985). Staphylococcal a-toxin was purified from culture media of Staphylococcus aureu8 Wood 46 strain according to Watanabe & Kato (1978). Numerical data are expressed as means + standard error.…”

Section: Introductionmentioning

confidence: 99%

Spontaneous release of nitric oxide inhibits electrical, Ca2+ and mechanical transients in canine gastric smooth muscle.

Ozaki

Blondfield

Hori

et al. 1992

The Journal of Physiology

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SUMMARY1. In canine antrum, rhythmic electrical activity consists of a rapid upstroke phase followed by a plateau depolarization. In response to slow waves, cytosolic Ca2" ([Ca2`],yt) and tension increased. 6. In muscles permeabilized by oc-toxin, cyclic GMP (100 /cM) and UV illumination inhibited Ca2`-induced contraction (at pCa 5 5).7. These data suggest that NO or NO-related compounds are spontaneously released in gastric muscles. These agents have two effects on excitation-contraction coupling: (i) inhibition (directly and/or indirectly) of the voltage-dependent Ca2" channels that participate in the plateau phase of slow waves, and (ii) reduction in the Ca21 sensitivity of the contractile element.

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Influence of Cys-130 S. aureus Alpha-toxin on Planar Lipid Bilayer and Erythrocyte Membranes

Krasilnikov

Yuldasheva

1997

Journal of Membrane Biology

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Replacement of an amino acid residue at position 130 -Gly by Cys- in the primary structure of Staphylococcus aureus alpha-toxin decreases the single-channel conductance induced by the toxin in planar lipid bilayers. Concomitantly, the pH value at which the channel becomes unable to discriminate between Cl- and K+ ions is also decreased. By contrast, the pH dependence of the efficiency of the mutant toxin to form ion channels in lipid bilayers was unchanged (maximum efficiency at pH 5.5-6.0). The asymmetry and nonlinearity of the current-voltage characteristics of the channel were increased by the point mutation but the diameter of the water pore induced by the mutant toxin, evaluated in lipid bilayers and in erythrocyte membranes, was found to be indistinguishable from that formed by wild-type toxin and equal to 2.4-2.6 nm. Alterations at the "trans mouth" were found to be responsible for all observed changes of the channel properties. This mouth is situated close to the surface of the second leaflet of a bilayer lipid membrane. The data obtained allows us to propose that the region around residue 130 in fact determines the main features of the ST-channel and takes part in the formation of the trans entrance of the channel.

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Purification and some properties of a lethal toxic fragment of staphylococcal α-toxin by tryptic digestion

Cited by 21 publications

References 19 publications

A Staphylococcal Α‐toxin Fragment

A Staphylococcal Α‐toxin Fragment

Spontaneous release of nitric oxide inhibits electrical, Ca2+ and mechanical transients in canine gastric smooth muscle.

Influence of Cys-130 S. aureus Alpha-toxin on Planar Lipid Bilayer and Erythrocyte Membranes

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