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通过转录组分析在秘鲁 Loxosceles laeta 蜘蛛毒液中鉴定的 astacin 样金属蛋白酶的多样性及其体外活性特征。

Diversity of astacin-like metalloproteases identified by transcriptomic analysis in Peruvian Loxosceles laeta spider venom and in vitro activity characterization.

机构信息

Biochemistry and Immunology Department, Federal University of Minas Gerais, Brazil; Genetic, Ecology and Evolution Department, Federal University of Minas Gerais, Brazil.

Ezequiel Dias Foundation, Minas Gerais, Brazil.

出版信息

Biochimie. 2019 Dec;167:81-92. doi: 10.1016/j.biochi.2019.08.017. Epub 2019 Aug 30.

Abstract

Loxosceles spiders are found in almost all countries of South America. In Peru, Loxosceles laeta species is the main responsible for the accidents caused by poisonous animals, being known as "killer spiders", due to the large number of fatal accidents observed. Astacin-like metalloproteases, named LALPs (Loxosceles astacin-like metalloproteases) are highly expressed in Loxosceles spiders venom gland. These proteases may be involved in hemorrhage and venom spreading, being relevant to the envenoming proccess. Thus, the aim of this work was to analyze Peruvian L. laeta venom gland transcripts using bioinformatics tools, focusing on LALPs. A cDNA library from Peruvian L. laeta venom glands was constructed and sequenced by MiSeq (Illumina) sequencer. After assembly, the resulting sequences were annotated, seeking out for similarity with previously described LALPs. Nine possible LALPs isoforms from Peruvian L. laeta venom were identified and the results were validated by in silico and in vitro experiments. This study contributes to a better understanding of the molecular diversity of Loxosceles venom and provide insights about the action of LALPs.

摘要

游走蛛遍布于南美洲的大部分国家。在秘鲁,拉氏游走蛛是引起动物毒性事故的主要元凶,因其造成的大量致命事故而被称为“杀人蜘蛛”。类似于 astacin 的金属蛋白酶,被命名为 LALPs(拉氏游走蛛类似 astacin 的金属蛋白酶),在游走蛛的毒腺中高度表达。这些蛋白酶可能参与了出血和毒液的扩散,与毒液的作用过程有关。因此,本工作旨在使用生物信息学工具分析秘鲁拉氏游走蛛的毒腺转录本,重点研究 LALPs。从秘鲁拉氏游走蛛的毒腺中构建了 cDNA 文库,并使用 MiSeq(Illumina)测序仪进行了测序。组装后,对得到的序列进行注释,寻找与先前描述的 LALPs 的相似性。从秘鲁拉氏游走蛛的毒液中鉴定出了 9 种可能的 LALPs 同工型,并通过计算机模拟和体外实验进行了验证。本研究有助于更好地了解拉氏游走蛛毒液的分子多样性,并深入了解 LALPs 的作用机制。

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