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比塞大泻湖中一种新型贝类生物毒素的纯化、毒性及功能特征分析。

Purification, Toxicity and Functional Characterization of a New Proteinaceous Mussel Biotoxin from Bizerte Lagoon.

机构信息

Institut Pasteur de Tunis, Laboratoire des Venins et Biomolécules Thérapeutiques, Université Tunis El Manar, 13 Place Pasteur, B.P. 74, 1002 Tunis-Belvédère, Tunisia.

CEA, Institut des Sciences du Vivant Frédéric Joliot, Département Médicaments et Technologies pour la Santé (DMTS), Service d'Ingénierie Moléculaire pour la Santé (SIMoS), Université Paris-Saclay, ERL 9004 CNRS/CEA, 91191 Gif sur Yvette, France.

出版信息

Toxins (Basel). 2020 Jul 30;12(8):487. doi: 10.3390/toxins12080487.

Abstract

The marine environment is known to be occupied by microorganisms. The potential toxicity of some of these marine microorganisms, that are capable of producing unknown biotoxins, has always been underestimated. Indeed, these biotoxins may be a threat to human health through the consumption of contaminated seafood and fish. For more than ten years, recurrent but atypical toxicity has been detected in mussels from Bizerte lagoon (North of Tunisia) during routine tests. In this study, we have isolated and characterized a new proteinaceous marine biotoxin, named Mussel Toxic Peptide (MTP). Using HPLC, electrophoresis and LC/MS studies, we showed that MTP has a protein characteristic UV-spectrum, can be visualized by protein specific reagents such as Coomassie, and has a molecular mass of 6.4 kDa. Patch-clamp experiments performed on cultured N18 neuroblastoma cells revealed that MTP (0.9-18 µM) markedly inhibited voltage-gated Na current, but was about 23 times less active in blocking voltage-gated K current at equimolar concentrations. To the best of our knowledge, this is the first time that a proteinaceous marine biotoxin with relatively high molecular mass is isolated and involved in the contamination of mussels harvested from shellfish farming areas.

摘要

海洋环境中存在着微生物。其中一些具有产生未知生物毒素能力的海洋微生物的潜在毒性一直被低估。事实上,这些生物毒素可能通过食用受污染的海鲜和鱼类对人类健康构成威胁。十多年来,在突尼斯北部比塞大泻湖的贻贝类常规检测中,一直检测到反复出现但非典型的毒性。在这项研究中,我们分离并鉴定了一种新型的蛋白类海洋生物毒素,命名为贻贝毒素肽(Mussel Toxic Peptide,MTP)。通过 HPLC、电泳和 LC/MS 研究,我们表明 MTP 具有蛋白质特有的紫外光谱,可以被考马斯亮蓝等蛋白质特异性试剂可视化,分子量为 6.4 kDa。在培养的 N18 神经母细胞瘤细胞上进行的膜片钳实验表明,MTP(0.9-18 μM)显著抑制电压门控 Na 电流,但在等摩尔浓度下,阻断电压门控 K 电流的活性约低 23 倍。据我们所知,这是首次分离出具有相对高分子质量的蛋白类海洋生物毒素,并参与了贝类养殖区贻贝类的污染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c915/7472388/f4068d6a430d/toxins-12-00487-g001.jpg

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