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菲律宾南部萨马尔眼镜蛇（Naja samarensis）的蛇毒蛋白质组学：短α-神经毒素作为主要致死成分，被菲律宾眼镜蛇抗蛇毒血清微弱交叉中和。

Snake Venom Proteomics of Samar Cobra (Naja samarensis) from the Southern Philippines: Short Alpha-Neurotoxins as the Dominant Lethal Component Weakly Cross-Neutralized by the Philippine Cobra Antivenom.

作者信息

Palasuberniam Praneetha, Chan Yi Wei, Tan Kae Yi, Tan Choo Hock

机构信息

Venom Research and Toxicology Laboratory, Department of Pharmacology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University Malaysia Sabah, Kota Kinabalu, Malaysia.

出版信息

Front Pharmacol. 2021 Dec 24;12:727756. doi: 10.3389/fphar.2021.727756. eCollection 2021.

DOI:10.3389/fphar.2021.727756
PMID:35002690
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8740184/
Abstract

The Samar Cobra, , is endemic to the southern Philippines and is a WHO-listed Category 1 venomous snake species of medical importance. Envenomation caused by results in neurotoxicity, while there is no species-specific antivenom available for its treatment. The composition and neutralization of venom remain largely unknown to date. This study thus aimed to investigate the venom proteome of for a comprehensive profiling of the venom composition, and to examine the immunorecognition as well as neutralization of its toxins by a hetero-specific antivenom. Applying C reverse-phase high-performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (LC-MS/MS), three-finger toxins (3FTx) were shown to dominate the venom proteome by 90.48% of total venom proteins. Other proteins in the venom comprised snake venom metalloproteinases, phospholipases A cysteine-rich secretory proteins, venom nerve growth factors, L-amino acid oxidases and vespryn, which were present at much lower abundances. Among all, short-chain alpha-neurotoxins (SαNTX) were the most highly expressed toxin within 3FTx family, constituting 65.87% of the total venom proteins. The SαNTX is the sole neurotoxic component of the venom and has an intravenous median lethal dose (LD) of 0.18 μg/g in mice. The high abundance and low LD support the potent lethal activity of venom. The hetero-specific antivenom, Philippine Cobra Antivenom (PCAV, raised against ) were immunoreactive toward the venom and its protein fractions, including the principal SαNTX. In efficacy study, PCAV was able to cross-neutralize the lethality of SαNTX albeit the effect was weak with a low potency of 0.20 mg/ml (defined as the amount of toxin completely neutralized per milliliter of the antivenom). With a volume of 5 ml, each vial of PCAV may cross-neutralize approximately 1 mg of the toxin . The findings support the potential para-specific use of PCAV in treating envenomation caused by while underscoring the need to improve the potency of its neutralization activity, especially against the highly lethal alpha-neurotoxins.

摘要

萨马尔眼镜蛇（学名：Naja samarensis），原产于菲律宾南部，是世界卫生组织列出的具有重要医学意义的1类毒蛇。其毒液会导致神经毒性，然而目前尚无针对其治疗的特异性抗蛇毒血清。迄今为止，萨马尔眼镜蛇毒液的成分及中和作用在很大程度上仍不为人知。因此，本研究旨在调查萨马尔眼镜蛇的毒液蛋白质组，以全面描绘毒液成分，并研究一种异种特异性抗蛇毒血清对其毒素的免疫识别及中和作用。通过反相高效液相色谱法（RP-HPLC）和串联质谱法（LC-MS/MS）分析，三指毒素（3FTx）在毒液蛋白质组中占主导地位，占毒液总蛋白的90.48%。毒液中的其他蛋白质包括蛇毒金属蛋白酶、磷脂酶A、富含半胱氨酸的分泌蛋白、毒液神经生长因子、L-氨基酸氧化酶和vespryn，其含量要低得多。其中，短链α-神经毒素（SαNTX）是3FTx家族中表达量最高的毒素，占毒液总蛋白的65.87%。SαNTX是毒液中唯一的神经毒性成分，在小鼠体内的静脉注射半数致死剂量（LD）为0.18μg/g。高丰度和低LD值表明萨马尔眼镜蛇毒液具有强大的致死活性。异种特异性抗蛇毒血清——菲律宾眼镜蛇抗蛇毒血清（PCAV，针对菲律宾眼镜蛇制备）对该毒液及其蛋白组分具有免疫反应性，包括主要的SαNTX。在疗效研究中，PCAV能够交叉中和SαNTX的致死性，尽管效果较弱，效价为0.20mg/ml（定义为每毫升抗蛇毒血清完全中和毒素的量）。每瓶5ml的PCAV大约可交叉中和1mg的毒素。这些发现支持了PCAV在治疗萨马尔眼镜蛇所致中毒方面的潜在异种特异性应用，同时强调需要提高其中和活性的效价，尤其是针对高度致死性的α-神经毒素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/b82cc46eb952/fphar-12-727756-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/aa0d18aff4a0/fphar-12-727756-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/16c1fd07476a/fphar-12-727756-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/f35f3159b536/fphar-12-727756-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/6851c8fe268d/fphar-12-727756-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/b82cc46eb952/fphar-12-727756-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/aa0d18aff4a0/fphar-12-727756-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/16c1fd07476a/fphar-12-727756-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/f35f3159b536/fphar-12-727756-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/6851c8fe268d/fphar-12-727756-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5135/8740184/b82cc46eb952/fphar-12-727756-g005.jpg

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本文引用的文献

[1]
A Quest for a Universal Plasma-Derived Antivenom Against All Elapid Neurotoxic Snake Venoms.

Front Immunol. 2021

[2]
Convergent evolution of pain-inducing defensive venom components in spitting cobras.

Science. 2021-1-22

[3]
A Neurotoxic Snake Venom without Phospholipase A: Proteomics and Cross-Neutralization of the Venom from Senegalese Cobra, (Subgenus: ).

Toxins (Basel). 2021-1-14

[4]
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Trans R Soc Trop Med Hyg. 2021-1-7

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Comparative venom proteomics of banded krait (Bungarus fasciatus) from five geographical locales: Correlation of venom lethality, immunoreactivity and antivenom neutralization.

Acta Trop. 2020-4-9

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Toxins (Basel). 2020-3-23

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Proteomic insights into short neurotoxin-driven, highly neurotoxic venom of Philippine cobra (Naja philippinensis) and toxicity correlation of cobra envenomation in Asia.

J Proteomics. 2019-6-12

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