The Fixation of Tetanus Toxin, Strychnine, Serotonin and other Substances by Ganglioside

Heyningen, W. E. van

doi:10.1099/00221287-31-3-375

Cited by 123 publications

(20 citation statements)

References 6 publications

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“…As described above, we have observed similar low levels of toxin-induced channel activity in asolectin membranes at a symmetric pH of 7.0. This activity was not affected by the presence of ganglioside GTlb (asolectin to ganglioside ratio 9:1), a putative receptor for tetanus toxin (23).…”

Section: Methodsmentioning

confidence: 95%

Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes.

Hoch¹,

Romero-Mira²,

Ehrlich³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

287

191

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The heavy chains of both botulinum neurotoxin type B and tetanus toxin form channels in planar bilayer membranes. These channels have pH-dependent and voltagedependent properties that are remarkably similar to those previously described for diphtheria toxin. Selectivity experiments with anions and cations show that the channels formed by the heavy chains of all three toxins are large; thus, these channels could serve as "tunnel proteins" for translocation of active peptide fragments. These findings support the hypothesis that the active fragments of botulinum neurotoxin and tetanus toxin, like that of diphtheria toxin, are translocated across the membranes of acidic vesicles.Diphtheria toxin (1), botulinum neurotoxin (2), and tetanus toxin (3) are proteins that are similar in origin and macrostructure. All three toxins are synthesized by bacteria (Corynebacterium diphtheriae, Clostridium botulinum, and Clostridium tetani) as single polypeptide chains (diphtheria toxin, "'60 kDa; clostridial neurotoxins, ==150 kDa). When exposed to trypsin or trypsin-like enzymes, they are cleaved to yield two-chain molecules in which a heavy-chain polypeptide is linked by a disulfide bond to a light-chain polypeptide. The two-chain structure is the active form of the three toxins.Various techniques have been used to generate polypeptide fragments from the toxins. The most straightforward of these is disulfide-bond reduction, which releases the heavy chain from the light chain. Alternatively, the clostridial neurotoxins have been exposed to limited proteolysis (e.g., with papain) to generate a fragment B and fragment C. Finally, traditional techniques have been used to select mutant organisms that synthesize incomplete toxins, such as the CRM45 fragment of diphtheria toxin, from which the B45 fragment can be formed. The various toxins and their fragments are illustrated in Fig. 1 Previous studies have shown that the amino terminus of the heavy chain from diphtheria toxin (18) and whole diphtheria toxin (19) form channels in lipid bilayers, and it has been proposed (18) that these channels provide the pathway for the light chain to cross membranes. Here we report that the heavy chains of both botulinum neurotoxin type B and tetanus toxin also form channels in lipid bilayers. Furthermore, for all three toxins, channel formation is maximal when the protein-containing (cis) side of the artificial membrane is at low pH (-4.0) and the opposite (trans) side is at pH -7.0, a pH gradient comparable to that across the membranes of acidic vesicles in cells. The channels for all three toxins are very large, as determined by selectivity experiments with large anions and cations, and this finding is compatible with the idea that the channels function as "tunnel proteins" for translocation of fully extended active fragments. In addition, tetanus toxin channels display a voltage dependence similar to that of diphtheria toxin channels, opening when positive voltages are applied to the cis side of artificial membranes and closing when nega...

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Section: Methodsmentioning

confidence: 95%

Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes.

Hoch¹,

Romero-Mira²,

Ehrlich³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

287

191

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“…Cells were grown on 0.1% gelatin-coated glass (for PCC7 cells) or tissue culture plastic coverslips (for C1003 cells). Stage-specific mouse embryonic antigen (SSEA-1) (24) and tetanus toxin binding sites (25) were revealed on cells fixed with 3% paraformaldehyde (10 min at room temperature). Mouse monoclonal anti-SSEA-1 antibody was generously provided by D. Solter (24).…”

Section: Methodsmentioning

confidence: 99%

Abundant expression of homeobox genes in mouse embryonal carcinoma cells correlates with chemically induced differentiation.

Deschamps¹,

Laaf²,

Joosen³

et al. 1987

Proc. Natl. Acad. Sci. U.S.A.

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Mammalian homeobox-containing genes might play a role in embryonal pattern formation. In favor of this view is the recently reported expression of such genes during mouse embryogenesis [Manley, J. L. & Levine, M. S. (1985) CeU 43, 1-2]. The embryo-derived stem cells and in particular the pluripotent embryonal carcinoma (EC) cell lines are generally considered as a valid model of early mouse development. Homeobox-containing genes were shown to be expressed in differentiating EC cells. We have analyzed the expression of several of these genes in three EC cell lines triggered to differentiate by alternative treatments in the presence or in the absence of retinoic acid. In both types of conditions, C17S1 (done 1003) and PCC7.S Aza R1 EC cells were induced to differentiate into mainly neurones, and PSA-1 EC cells were induced to differentiate into a large spectrum of tissue derivatives. Induction to high levels of expression of several homeobox-containing genes during differentiation occurs only in the presence of retinoic acid. Nonchemical treatment triggering differentiation does not lead to detectable expression of these genes. Accumulation to high amounts of homeobox-containing gene transcripts in these experiments seems to correlate with retinoic acid-induced EC cell differentiation rather than with EC cell differentiation as such.

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“…The precise nature of the 5-HT receptor has been a subject of interest for several years. Woolley and Gommni (51) (52) demonstrated that gangliosides in aqueous media could bind 5-HT as well as other pharmacologic agents such as lysergic acid diethylamide and chlorpromazine. Gielen Human Platelet Gangliosides (53,54) (35) reported that 74% of the total sialic acid of frozen and thawed platelets was liberated by neuraminidase.…”

Section: Discussionmentioning

confidence: 99%

Studies on Human Platelet Gangliosides

Marcus¹,

Ullman²,

Safier³

1972

J. Clin. Invest.

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A B S T R A C T Gangliosides, glycosphingolipids which contain sialic acid, were studied in human platelets. They represented 0.5% of the platelet lipids and accounted for 6% of the total neuraminic acid content of platelets. Three major ganglioside fractions were identified and characterized. Ganglioside I was hematoside (G6) and comprised 92% of the platelet gangliosides. It contained glucose, galactose, and sialic acid in molar ratios of 1: 1: 1 and no hexosamnine. The imiajor fatty acid was behenate (22: 0). Ganglioside I w-as also identified in isolated platelet granules and membranes. Ganglioside II (5%) contained glucose, galactose. sialic acid, and hexosamines (molar ratios 1: 2: 1: 1). The hexosamines were glucosamine (72%) and galactosamine (28%). It was therefore designated as ganglioside lacto-N-neotetraose. Ganglioside III (2%) contained disialosyllactosyl ceramide (G3A) as well as two other gangliosides which could not be precisely characterized. Gangliosides I, II, and III were susceptible to the action of Clostridiuin perfringens neuraminidase as evidenced by full recovery of sialic acid in its free form after incubation. Neutral platelet glycolipids were qualitatively examined by thinlayer chromatography. The major component was lactosvI ceramide.Interactions of gangliosides I and III and serotonin-'4Cwere examined in an equilibrium dialysis system at 4°C. The gangliosides bound serotonin-'4C in relatively small quantities, wlhereas cointrol lipids w-ere negative. The binding was essentially tinchainged by reverse dialysis, ultracentrifugation anld stubsequen-t tlhin-laver chromatography. The results are comparable to the previously observed nonmetabolic interactions between wlhole platelets and serotonin in the cold. It is suggested that the orientation and specific distribtution of platelet membrane glycolipids may be important determinants of the unique surface properties of platelets.

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The Fixation of Tetanus Toxin, Strychnine, Serotonin and other Substances by Ganglioside

Cited by 123 publications

References 6 publications

Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes.

Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes.

Abundant expression of homeobox genes in mouse embryonal carcinoma cells correlates with chemically induced differentiation.

Studies on Human Platelet Gangliosides

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