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Toxicon ; 99: 95-101, 2015 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-25817004

RESUMO

Development of novel analytical tools to detect marine biotoxins has been warranted in view of the apparent global pervasiveness of algal-derived shellfish poisoning, and the limitations of existing methods. Here, we describe the initial phase in the development and evaluation of a tyrosine-containing analog of µ-conotoxin (µ-CTX) GIIIA as an alternative to saxitoxin (STX) in a receptor binding assay (RBA) for paralytic shellfish poisons. The peptide analog was synthesized and characterized for structure and bioactivity. The major product of oxidation elicited paralytic symptoms in mice at a minimum dose of 1.31 mg kg(-1) (i.p.). Mass spectrometry analysis of the bioactive peptide gave a molecular mass of 2637.52 Da that was close to the predicted value. Iodination via chloramine-T produced non-, mono- and di-iodinated peptides (respectively, NIP, MIP and DIP). Competition assays against (3)H-STX revealed higher Ki and EC50 (P < 0.0001, ANOVA) indicating reduced affinity for the receptor, and limited displacement of receptor-bound STX. However, subsequent use of MIP may extend the application of RBA to detect small changes in toxin levels owing to its likely enhanced displacement by STX. This may be useful in analyzing samples with toxicities near the regulatory limit, or in establishing baseline values in high risk environments.


Assuntos
Conotoxinas/análise , Inspeção de Alimentos/métodos , Proteínas Musculares/metabolismo , Neurotoxinas/análise , Saxitoxina/análise , Substituição de Aminoácidos , Animais , Ligação Competitiva , Bioensaio , Conotoxinas/química , Conotoxinas/metabolismo , Conotoxinas/toxicidade , Contaminação de Alimentos , Halogenação , Humanos , Ligantes , Masculino , Camundongos , Camundongos Endogâmicos ICR , Neurotoxinas/química , Neurotoxinas/metabolismo , Neurotoxinas/toxicidade , Peptídeos/análise , Peptídeos/química , Peptídeos/metabolismo , Peptídeos/toxicidade , Filipinas , Ratos Sprague-Dawley , Saxitoxina/metabolismo , Saxitoxina/toxicidade , Frutos do Mar/análise , Intoxicação por Frutos do Mar/etiologia , Intoxicação por Frutos do Mar/metabolismo , Trítio
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