Toxicon : official journal of the International Society on Toxinology · 2026
Omics-guided discovery and functional profiling of Da-29, a bioactive serine protease from Deinagkistrodon acutus venom, with antithrombotic activity.
СтатьяНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Chen Caojuan, Ding Peng, Tao Xinru, Li Jingxi, Liu Pingan, Cai Yuan, Peng Yanmei
Аннотация
BACKGROUND: Snake venom serine proteases (SVSPs) are critical toxic components that interfere with the hemostatic balance and serve as a key candidate library for anticoagulant drug development. Despite the diversity of SVSPs in Deinagkistrodon acutus venom, systematic studies of low-abundance SVSPs with thrombin-like activity profiles are limited. Hence, this research aimed to isolate and characterize a bioactive SVSP candidate from D. acutus venom using integrated multi-omics and to assess its antithrombotic effects and bleeding risk.
METHODS: PacBio full-length transcriptome sequencing combined with label-free quantitative proteomics was used to screen SVSP candidates with matched transcription-translation associations. The target protein Da-29 was purified using gel filtration and reverse-phase chromatography, and its primary structure was confirmed using transcriptome sequencing and liquid chromatography-tandem mass spectrometry. In vitro enzymatic activity and fibrinogen hydrolysis specificity, as well as in vivo antithrombotic efficacy and bleeding risk, were evaluated using rodent thrombosis and bleeding models.
RESULTS: Da-29 was observed to be a 260-amino-acid SVSP containing a conserved catalytic triad and an S1 substrate-binding pocket. It exhibited significant amidase and fibrinogen-clotting activities and selectively hydrolyzed fibrinogen Aα and Bβ chains. Furthermore, Da-29 exerted dose-associated antithrombotic effects in vivo, extending the coagulation time, reducing the fibrinogen/TXB2 levels, and upregulating 6-keto-PGF1α, without causing considerable bleeding risks at effective doses.
CONCLUSION: Da-29 exhibits antithrombotic activity associated with fibrinogen degradation and linked to concurrent alterations in TXB2 and 6-keto-PGF1α levels, establishing its potential as an antithrombotic experimental lead. This integrated omics strategy can effectively identify low- abundance bioactive molecules from snake venom, offering a reproducible paradigm for the discovery of functional proteins in natural products.
METHODS: PacBio full-length transcriptome sequencing combined with label-free quantitative proteomics was used to screen SVSP candidates with matched transcription-translation associations. The target protein Da-29 was purified using gel filtration and reverse-phase chromatography, and its primary structure was confirmed using transcriptome sequencing and liquid chromatography-tandem mass spectrometry. In vitro enzymatic activity and fibrinogen hydrolysis specificity, as well as in vivo antithrombotic efficacy and bleeding risk, were evaluated using rodent thrombosis and bleeding models.
RESULTS: Da-29 was observed to be a 260-amino-acid SVSP containing a conserved catalytic triad and an S1 substrate-binding pocket. It exhibited significant amidase and fibrinogen-clotting activities and selectively hydrolyzed fibrinogen Aα and Bβ chains. Furthermore, Da-29 exerted dose-associated antithrombotic effects in vivo, extending the coagulation time, reducing the fibrinogen/TXB2 levels, and upregulating 6-keto-PGF1α, without causing considerable bleeding risks at effective doses.
CONCLUSION: Da-29 exhibits antithrombotic activity associated with fibrinogen degradation and linked to concurrent alterations in TXB2 and 6-keto-PGF1α levels, establishing its potential as an antithrombotic experimental lead. This integrated omics strategy can effectively identify low- abundance bioactive molecules from snake venom, offering a reproducible paradigm for the discovery of functional proteins in natural products.