Clostridium botulinum type A haemagglutinin-positive progenitor toxin (HA+-PTX) binds to oligosaccharides containing Galβ1-4GlcNAc through one subcomponent of haemagglutinin (HA1)

Inoue, Kaoru; Fujinaga, Yukako; Honke, Koichi; Arimitsu, Hideyuki; Mahmut, Nazira; Sakaguchi, Yoshihiko; Ohyama, Tohru; Inoue, Katsuhiro; Oguma, Keiji

doi:10.1099/00221287-147-4-811

Cited by 49 publications

(28 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HA proteins appear to play an important role in absorption of PTC by interacting with oligosaccharides on intestinal epithelial cells (Fujinaga et al 1997;Fujinaga et al 2004;Inoue et al 2001;Kojima et al 2005;Nakamura et al 2007) (also see chapter 3). One Nacetylneuraminic acid (Neu5Ac) binding site has been reported in HA-70b of BoNT/C, which is located in a cleft formed by domains II and III (Nakamura et al 2009).…”

Section: Has Mediate Toxin-host Interactionsmentioning

confidence: 99%

Assembly and Function of the Botulinum Neurotoxin Progenitor Complex

Gu¹,

Jin²

2012

Current Topics in Microbiology and Immunology

View full text Add to dashboard Cite

Botulinum neurotoxins (BoNTs) are among the most poisonous substances known to man, but paradoxically, BoNT-containing medicines and cosmetics have been used with great success in the clinic. Accidental BoNT poisoning mainly occurs through oral ingestion of food contaminated with Clostridium botulinum. BoNTs are naturally produced in the form of progenitor toxin complexes, which are high molecular weight (up to ~900 kDa) multi-protein complexes composed of BoNT and several non-toxic neurotoxin-associated proteins (NAPs). NAPs protect the inherently fragile BoNTs against the hostile environment of the gastrointestinal tract and help BoNTs pass through the intestinal epithelial barrier before they are released into the general circulation. These events are essential for ingested BoNTs to gain access to motoneurons, where they inhibit neurotransmitter release and cause muscle paralysis. In this review, we discuss the structural basis for assembly of NAPs and BoNT into the progenitor toxin complexes that protect BoNT and facilitate its delivery into the bloodstream.

show abstract

Section: Has Mediate Toxin-host Interactionsmentioning

confidence: 99%

Assembly and Function of the Botulinum Neurotoxin Progenitor Complex

Gu¹,

Jin²

2012

Current Topics in Microbiology and Immunology

View full text Add to dashboard Cite

show abstract

“…Only the 16 S toxin bound obviously to the epithelial cells and time to death after injection with 16 S toxin was measurably shorter compared with 12 S toxin or neurotoxin [13]. Furthermore, in a way comparable to 16 S toxin, recombinant proteins of HA1 and HA3 could bind both epithelial cells and erythrocytes [14,15]. Some monoclonal antibodies against HA1 reduced the 16 S toxin binding to the epithelial cells and neutralised the oral toxicity of small amounts of 16 S toxin [16].…”

Section: Introductionmentioning

confidence: 99%

Mucosal immunisation with Clostridium botulinum type C 16 S toxoid and its non-toxic component

Mahmut¹,

Inoue

Fujinaga³

et al. 2002

Journal of Medical Microbiology

Self Cite

View full text Add to dashboard Cite

Clostridium botulinum types C and D produce a 16 S (500 kDa) toxin that is formed by conjugation of neurotoxin with a non-toxic component (nonTox). The amino acid sequences of type C and D nonTox components are almost identical. In a previous report it was proposed that nonTox is necessary for the effective absorption of the toxin from the small intestine. This suggested the hypothesis that mucosal immunity against nonTox in the small intestine might prevent the absorption of both C-and D-16 S toxins. The nonTox was purified from a mutant strain, (C)-N71, that does not produce neurotoxin. This nonTox or detoxified C-16 S toxin were mixed with adjuvant (a mutant form of heat-labile toxin of Escherichia coli), and inoculated into mice via the nasal or oral route, or both. The mice inoculated nasally four times with nonTox or toxoid produced high levels of antibodies (including IgA) against the immunogens, both in intestinal fluids and sera. When these nonTox-immunised mice were challenged orally with 2 and 20 oral minimum lethal doses (MLD) of C-or D-16 S toxins, the same results were obtained with both C and D; the mice survived after challenge with 2 MLD of either C or D but were killed by 20 MLD of either toxin although the time to death was significantly longer than in the control non-immunised mice. These results indicate that the local anti-nonTox antibodies reduce absorption of both C-and D-16 S toxins from the small intestine. The C-16 S toxoid-immunised mice showed similar behaviour with type D toxin challenge, probably due to the same mechanism, but were protected against 20 MLD of C-16 S toxin.

show abstract

“…The toxin titer (minimum lethal dose [MLD]/ml) was obtained by injecting the diluted preparation into three mice intraperitoneally, each receiving 0.5 ml. The HA titer was obtained by reacting twofold-diluted preparations with an equal volume of 1% neuraminidase-treated human erythrocytes in a microtiter plate (7,8). Peak 3 possessed both toxicity (8 ϫ 10 7 MLD/ml) and HA activity.…”

mentioning

confidence: 99%

“…Recently, it was found that serotype A and B HA-positive toxins could bind to both erythrocytes and the epithelial cells of the small intestine mainly via HA1 and that these bindings were effectively inhibited by lactose (3,8) (data concerning serotype B have not been published yet), indicating that HApositive toxins can bind to lactose via HA1. Based on these data, we planned to separate HA-positive 16S toxin and HAnegative 12S toxin by use of affinity gel column-linked lactose; it was speculated that the 12S toxin would pass through the showed both a high HA titer and toxicity, indicating that the former is 12S toxin and the latter is 16S toxin (Fig.…”

mentioning

confidence: 99%

Purification of Fully ActivatedClostridium botulinumSerotype B Toxin for Treatment of Patients with Dystonia

et al. 2003

Self Cite

View full text Add to dashboard Cite

Clostridium botulinum serotype B toxins 12S and 16S were separated by using a beta-lactose gel column at pH 6.0; toxin 12S passed through the column, whereas toxin 16S bound to the column and eluted with lactose. The fully activated neurotoxin was obtained by applying the trypsin-treated 16S toxin on the same column at pH 8.0; the neurotoxin passed through the column, whereas remaining nontoxic components bound to the column. The toxicity of this purified fully activated neurotoxin was retained for a long period by addition of albumin in the preparation.Clostridium botulinum strains produce immunologically distinct neurotoxins (serotypes A to G). The molecular masses of neurotoxin types A to G are approximately 150 kDa. The neurotoxins are produced as a single form and become dichain-form light (50-kDa) and heavy (100-kDa) chains by cleavage with proteases such as trypsin at about one-third of the distance from the amino terminus, and the toxic activity of the dichain form becomes fully activated (1). In culture fluid and food with acidic conditions, the neurotoxins associate with nontoxic components and form large complexes designated progenitor toxins. Under alkaline conditions, the progenitor toxins dissociate into neurotoxin and nontoxic components (10, 18). The progenitor toxins are found in three forms with molecular masses of 900 kDa (19S), 500 kDa (16S), and 300 kDa (12S) (14). The 12S toxin is composed of a neurotoxin and a nontoxic component having no hemagglutinin (HA) activity (designated nontoxic non-HA [NTNH]), whereas the 16S and 19S toxins are composed of a neurotoxin, NTNH, and HA. The serotype A strain produces three forms of toxins (19S, 16S, and 12S). Type B, C, and D strains produce the 16S and the 12S toxins. We purified different-sized progenitor toxins from serotype A, C, and D cultures (2,4,6,12,13) and demonstrated that (i) the 19S toxin is a dimer of the 16S toxin; (ii) HA consists of four subcomponents with molecular masses of 52 to 53, 33 to 35, 19 to 23, and 15 to 17 kDa, designated here as HA3b, HA1, HA3a, and HA2, respectively; and (iii) NTNH of the 12S toxin has a cleavage site(s) at the N-terminal region.Recently, serotype A and B progenitor toxins have been used for treating patients with strabismus, blepharospasm, nystagmus, facial spasm, spastic aphonia, and many other forms of dystonia (9, 11). In both toxin types, progenitor toxins are used because they are easily obtained and are more stable than neurotoxin. The treatment is very effective but has a serious side effect for some patients in whom antiprogenitor toxins, including antineurotoxin antibodies, are produced after several injections. It seems that using neurotoxin alone is better than using the progenitor toxin (a complex of the neurotoxin and a nontoxic component). Furthermore, it has been reported that serotype B toxin, which is used therapeutically at present, is partially cleaved (16) and therefore the toxin is not fully activated. In this paper, we report a simple procedure for largescale purification of botu...

show abstract

Clostridium botulinum type A haemagglutinin-positive progenitor toxin (HA+-PTX) binds to oligosaccharides containing Galβ1-4GlcNAc through one subcomponent of haemagglutinin (HA1)

Abstract: Haemagglutinin (HA) activity of

Cited by 49 publications

References 28 publications

Assembly and Function of the Botulinum Neurotoxin Progenitor Complex

Assembly and Function of the Botulinum Neurotoxin Progenitor Complex

Mucosal immunisation with Clostridium botulinum type C 16 S toxoid and its non-toxic component

Purification of Fully ActivatedClostridium botulinumSerotype B Toxin for Treatment of Patients with Dystonia

Contact Info

Product

Resources

About