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Determining the success of eradication of an invasive species requires a way to decide when its risk of reoccurrence has become acceptably low. In Japan, the area populated by the Argentine ant, Linepithema humile (Mayr), is expanding, and eradication via chemical treatment is ongoing at various locations. One such program in Tokyo was apparently successful, because the ant population decreased to undetectable levels within a short time. However, construction of a population model for management purposes was difficult because the probability of detecting ants decreases rapidly as the population collapses. To predict the time when the ant was eradicated, we developed a multinomial-mixture model for chemical eradication based on monthly trapping data and the history of pesticide applications. We decided when to declare that eradication had been successful by considering both 'eradication' times, which we associated with eradication probabilities of 95% and 99%, and an optimal stopping time based on a 'minimum expected economic cost' that considered the possibility that surveys were stopped too soon. By applying these criteria, we retroactively declared that Argentine ants had been eradicated 38-42 months after the start of treatments (16-17 months after the last sighting).
Assuntos
Formigas/crescimento & desenvolvimento , Teorema de Bayes , Entomologia/métodos , Controle de Insetos/métodos , Inseticidas/toxicidade , Espécies Introduzidas/estatística & dados numéricos , Modelos Estatísticos , Animais , Formigas/efeitos dos fármacosRESUMO
A novel antimicrobial peptide, anoplin, was purified from the venom of the solitary wasp Anoplius samariensis. The sequence was mostly analyzed by mass spectrometry, which was corroborated by solid-phase synthesis. Anoplin, composed of 10 amino acid residues, Gly-Leu-Leu-Lys-Arg-Ile-Lys-Thr-Leu-Leu-NH2, has a high homology to crabrolin and mastoparan-X, the mast cell degranulating peptides from social wasp venoms, and, therefore, can be predicted to adopt an amphipathic alpha-helix secondary structure. In fact, the circular dichroism (CD) spectra of anoplin in the presence of trifluoroethanol or sodium dodecyl sulfate showed a high content, up to 55%, of the alpha-helical conformation. A modeling study of anoplin based on its homology to mastoparan-X supported the CD results. Biological evaluation using the synthetic peptide revealed that this peptide exhibited potent activity in stimulating degranulation from rat peritoneal mast cells and broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria. Therefore, this is the first antimicrobial component to be found in the solitary wasp venom and it may play a key role in preventing potential infection by microorganisms during prey consumption by their larvae. Moreover, this peptide is the smallest among the linear alpha-helical antimicrobial peptides hitherto found in nature, which is advantageous for chemical manipulation and medical application.
Assuntos
Antibacterianos/isolamento & purificação , Oligopeptídeos/química , Oligopeptídeos/isolamento & purificação , Venenos de Vespas/química , Venenos de Vespas/isolamento & purificação , Sequência de Aminoácidos , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos , Degranulação Celular , Cromatografia Líquida de Alta Pressão , Dicroísmo Circular , Feminino , Mastócitos/efeitos dos fármacos , Mastócitos/fisiologia , Testes de Sensibilidade Microbiana , Modelos Moleculares , Oligopeptídeos/farmacologia , Ratos , Alinhamento de Sequência , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Venenos de Vespas/farmacologia , VespasRESUMO
Micro-scale (sub-pmol) isolation and sequence determination of three peptides from the venom of the solitary spider wasp Cyphononyx dorsalis is described. We isolated two novel peptides Cd-125 and Cd-146 and a known peptide Thr(6)-bradykinin from only two venom sacs of solitary spider wasp Cyphononyx dorsalis without bioassay-guided fractionation, but instead guided by MALDI-TOF MS. The MALDI-TOF MS analysis of each fraction showed the purity and molecular weight of the components, which led to the isolation of the peptides virtually without loss of sample amount. The sequences of the novel peptides Cd-125 (Asp-Thr-Ala-Arg-Leu-Lys-Trp-His) and Cd-146 (Ser-Glu-Thr-Gly-Asn-Thr-Val-Thr-Val-Lys-Gly-Phe-Ser-Pro-Leu-Arg) were determined by Edman degradation together with mass spectrometry, and finally corroborated by solid-phase synthesis. The known peptide Thr(6)-bradykinin (Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg) was identified by comparison with the synthetic authentic specimen. This is the first example for any kinins to be found in Pompilidae wasp venoms. The procedure reported here can be applicable to studies on many other components of solitary wasp venoms with limited sample availability.
Assuntos
Venenos de Vespas/química , Sequência de Aminoácidos , Animais , Feminino , Espectrometria de Massas , Dados de Sequência Molecular , Venenos de Vespas/isolamento & purificaçãoRESUMO
The structural specificity of alpha-PMTX, a novel peptide toxin derived from wasp venom has been studied on the neuromuscular synapse in the walking leg of the lobster. alpha-PMTX is known to induce repetitive action potentials in the presynaptic axon due to sodium channel inactivation. We synthesized 29 analogs of alpha-PMTX by substituting one or two amino acids and compared threshold concentrations of these mutant toxins for inducing repetitive action potentials. In 13 amino acid residues of alpha-PMTX, Arg-1, Lys-3 and Lys-12 regulate the toxic activity because substitution of these basic amino acid residues with other amino acid residues greatly changed the potency. Determining the structure-activity relationships of PMTXs will help clarifying the molecular mechanism of sodium channel inactivation.
Assuntos
Junção Neuromuscular/fisiologia , Neurotoxinas/metabolismo , Canais de Sódio/metabolismo , Transmissão Sináptica/fisiologia , Venenos de Vespas/metabolismo , Substituição de Aminoácidos/genética , Animais , Proteínas de Insetos , Nephropidae , Junção Neuromuscular/efeitos dos fármacos , Neurotoxinas/genética , Neurotoxinas/toxicidade , Ligação Proteica , Relação Estrutura-Atividade , Transmissão Sináptica/efeitos dos fármacos , Venenos de Vespas/genética , Venenos de Vespas/toxicidadeRESUMO
A new mast cell degranulating peptide, eumenine mastoparan-AF (EMP-AF), was isolated from the venom of the solitary wasp Anterhynchium flavomarginatum micado, the most common eumenine wasp found in Japan. The structure was analyzed by FAB-MS/MS together with Edman degradation, which was corroborated by solid-phase synthesis. The sequence of EMP-AF, Ile-Asn-Leu-Leu-Lys-Ile-Ala-Lys-Gly-Ile-Ile-Lys-Ser-Leu-NH(2), was similar to that of mastoparan, a mast cell degranulating peptide from a hornet venom; tetradecapeptide with C-terminus amidated and rich in hydrophobic and basic amino acids. In fact, EMP-AF exhibited similar activity to mastoparan in stimulating degranulation from rat peritoneal mast cells and RBL-2H3 cells. It also showed significant hemolytic activity in human erythrocytes. Therefore, this is the first example that a mast cell degranulating peptide is found in the solitary wasp venom. Besides the degranulation and hemolytic activity, EMP-AF also affects on neuromuscular transmission in the lobster walking leg preparation. Three analogs EMP-AF-1 approximately 3 were snythesized and biologically tested together with EMP-AF, resulting in the importance of the C-terminal amide structure for biological activities.
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Degranulação Celular/efeitos dos fármacos , Mastócitos/efeitos dos fármacos , Venenos de Vespas/química , Venenos de Vespas/farmacologia , Sequência de Aminoácidos , Animais , Cromatografia Líquida de Alta Pressão , Feminino , Hemólise/efeitos dos fármacos , Humanos , Espectrometria de Massas , Dados de Sequência Molecular , Nephropidae , Conformação Proteica , Ratos , Ratos Sprague-Dawley , Espectrofotometria Ultravioleta , Venenos de Vespas/isolamento & purificaçãoRESUMO
The use of mass spectrometry, in which high-energy CID and charge remote fragmentation both of protonated and sodium-attached molecular ions was applied, afforded the structural elucidation of a new acylpolyaminetoxin with Mw=801 Da from the venom of the Brazilian garden spider Nephilengys cruentata. In spite of having the same Mw of the NPTX-2, previously described in the venom of the Joro spider Nephila clavata, neither toxins are isomers. In order to differentiate them by using the most usual nomenclature, the new toxin was named NPTX-801C and the NPTX-2 was renamed to NPTX-801E. Both toxins have as common structure the 4-hydroxyindole-3-acetyl-asparaginyl-cadaveryl moiety in their molecules and their structure may be represented in a simplified way: NPTX-801E is HO-indole-Asn-Cad-Pta-Orn-Arg and NPTX-801C is HO-indole-Asn-Cad-Gly-Put-Pta-Pta.
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Poliaminas/isolamento & purificação , Venenos de Aranha/química , Aminoácidos/análise , Animais , Brasil , Cromatografia Líquida de Alta Pressão , Peso Molecular , Poliaminas/química , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Espectrofotometria Ultravioleta , Venenos de Aranha/isolamento & purificaçãoRESUMO
A new strategy to characterize glutaminergic blocker acylpolymines stored in a spider venom with mass spectrometry is described. The crude spider venom extracts are amenable to direct MALDI mass spectrometry analysis which provides a rapid and accurate means of measuring the molecular weights of acylpolyamines without the isolation of individual samples. Compared with the previously developed mu-column HPLC/MS method, this procedure provides more efficient detection and identification of complex venom constituents. Twenty-five acylpolyamines were detected from Brazilian garden spider Nephilengys cruentata crude venom extracts by both HPLC/MS and MALDI-mass spectrometry. These acylpolyamine structures were determined by high-energy collision induced dissociation MS/MS method. Most of the compounds were classified into the previously reported generalized structures types A to D, which were found from the venom of Nephilengys borbonica. The structures of four acylpolyamines (M + H)+, m/z 623, 646, 688, and 745, which were not contained in the venom of Nephilengys borbonicare were determined to have arginine at the polyamine chain terminal and were named NPTX-622, -645, -687, and -744, respectively.