Dataset Information

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2).

ABSTRACT: Background:Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors. Methods:The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed. Results:V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (?/?)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4. Conclusion:The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.

SUBMITTER: Rungsa P

PROVIDER: S-EPMC6892566 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Heterologous expression and mutagenesis of recombinant <i>Vespa affinis</i> hyaluronidase protein (rVesA2).

Rungsa Prapenpuksiri P Janpan Piyapon P Saengkun Yutthakan Y Jangpromma Nisachon N Klaynongsruang Sompong S Patramanon Rina R Uawonggul Nunthawun N Daduang Jureerut J Daduang Sakda S

The journal of venomous animals and toxins including tropical diseases 20191205

<h4>Background</h4>Crude venom of the banded tiger wasp<i>Vespa affinis</i> contains a variety of enzymes including hyaluronidases, commonly known as spreading factors.<h4>Methods</h4>The cDNA cloning, sequence analysis and structural modelling of <i>V. affinis</i> venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed.<h4>Results</h4><i>V. affinis</i> venom hyaluronidase full sequence is composed of 331 amino acids, with fou ...[more]

PMID: 31839801

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Project description:Distinct animal lineages have convergently recruited venoms as weaponry for prey capture, anti-predator defence, conspecific competition, or a combination thereof. Most studies, however, have been primarily confined to a narrow taxonomic breadth. The venoms of cone snails, snakes, spiders and scorpions remain particularly well-investigated. Much less explored are the venoms of wasps (Order: Hymenoptera) that are infamous for causing excruciating and throbbing pain, justifying their apex position on Schmidt's pain index, including some that are rated four on four. For example, the lesser banded wasp (V. affinis) is clinically important yet has only been the subject of a few studies, despite being commonly found across tropical and subtropical Asia. Stings from these wasps, especially from multiple individuals of a nest, often lead to clinically severe manifestations, including mastocytosis, myasthenia gravis, optic neuropathy, and life-threatening pathologies such as myocardial infarction and organ failure. However, their venom composition and activity remain unexplored in the Indian subcontinent. Here, we report the proteomic composition, transcriptomic profile, and biochemical and pharmacological activities of V. affinis venom from southern India. Our findings suggest that wasp venoms are rich in diverse toxins that facilitate antipredator defence. Biochemical and pharmacological assessments reveal that these toxins can exhibit significantly higher activities than their homologues in medically important snakes. Their ability to exert potent effects on diverse molecular targets makes them a treasure trove for discovering life-saving therapeutics. Fascinatingly, wasp venoms, being evolutionarily ancient, exhibit a greater degree of compositional and sequence conservation across very distant populations/species, which contrasts with the patterns of venom evolution observed in evolutionarily younger lineages, such as advanced snakes and cone snails.

Dataset Information

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2).

Publications

Heterologous expression and mutagenesis of recombinant <i>Vespa affinis</i> hyaluronidase protein (rVesA2).

Similar Datasets

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