Analysis of the Composition of Deinagkistrodon acutus Snake Venom Based on Proteomics, and Its Antithrombotic Activity and Toxicity Studies
Abstract
:1. Introduction
2. Material and Methods
2.1. Materials Preparation
2.2. Venom Proteome Analysis
2.2.1. Nano-Liquid Chromatography–Tandem Mass Spectrometry Analysis
2.2.2. Protein Identification
2.2.3. Protein Function and Interaction Analyses
2.3. Lethal Activity of Deinagkistrodon acutus Venom
2.4. In Vitro Platelet Aggregation Assay
2.5. Anti-Coagulant Activity
Blood Clotting Assays
2.6. Collagen-Adrenalin-Induced Acute Pulmonary Thrombosis in Mice
2.7. Carrageenan-Induced Tail Thrombosis in Mice
2.8. Flow Restriction of the Inferior Vena Cava (IVC) in Mouse Induced Thrombosis
2.9. Statistical Analysis
3. Results
3.1. Proteomic Analysis of the Venom of Deinagkistrodon Acutus
3.2. Acute Toxicity Test of Deinagkistrodon acutus Venom
3.3. Coagulant Activity In Vivo
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
ANOVA | Analysis of variance |
ADP | Adenosine diphosphate |
PLT | Platelet |
PRP | Platelet Rich Plasma |
PPP | Platelet-Poor Plasma |
SPF | Specific pathogen Free |
IVC | Inferior Vena Cava |
LC-MS/MS | Liquid chromatography-tandem mass spectrometer |
GO | Gene ontology Terms |
iBAQ | Intensity based absolute quantification |
References
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Protein ID | Description | Coverage | Protein Mass | Peptide | Unique Peptide | Uniq Spectrum | snak_iBAQ | % | Function |
---|---|---|---|---|---|---|---|---|---|
CAJ85790.1 | Phospholipase A2, acidic precursor, partial [Deinagkistrodon acutus] | 0.9756 | 14,031.6 | 25 | 25 | 1191 | 6,623,486.35 | 13.80 | Binding catalytic_activity, Inhibits platelet aggregation. |
sp|Q9DEA2.1|SLCA_DEIAC | Snaclec agkicetin-C subunit alpha; | 0.7935 | 17,798.4 | 30 | 9 | 259 | 6,026,939.81 | 12.56 | Binding, Inhibits platelet aggregation, Inhibits platelet adhesion. |
AAM22784.1 | Antithrombin A B-chain [Deinagkistrodon acutus] | 0.7671 | 16,688 | 22 | 22 | 678 | 5,987,824.836 | 12.48 | Inhibits platelet aggregation, Inhibits platelet adhesion. |
AAL36975.1 | Lys-49 phospholipase A2 precursor [Deinagkistrodon acutus] | 0.6739 | 15,803.4 | 35 | 35 | 550 | 5,154,993.67 | 10.74 | Binding catalytic_activity, Indirect hemolytic activity and anticoagulant activity |
AAF76377.1 | Thrombin-like protein acutobin precursor [Deinagkistrodon acutus] | 0.6923 | 28,814.8 | 29 | 25 | 917 | 3,435,511.45 | 7.16 | Catalytic_activity, Coagulates human fibrinogen by hydrolysis of the alpha chains. |
SPQ1ZY03.1|PA2B_DEIAC | Basic phospholipase A2 DAV-N6; Short = svPLA2; AltName: Full = Phosphatidylcholine 2-acylhydrolase; Flags: Precursor | 0.7174 | 15,853.5 | 16 | 16 | 310 | 2,603,255.33 | 5.43 | Binding catalytic_activity, Blocking acetylcholine release from the nerve termini. |
AAL66390.1 | Antithrombin 1 B chain [Deinagkistrodon acutus] | 0.8591 | 17,249.8 | 41 | 15 | 274 | 2,102,366.73 | 4.38 | Inhibits platelet aggregation, Inhibits platelet adhesion. |
SP|Q9W7S2.1|VM11_DEIAC | Snake venom metalloproteinase aculysin-1; Short = SVMP; Flags: Precursor | 0.4604 | 47,304.2 | 105 | 58 | 1689 | 1,918,894.51 | 4.00 | Binding catalytic_activity, Hemorrhagic activity |
SP|Q9DEF9.1|SLAA_DEIAC | Snaclec anticoagulant protein subunit A; Short = AaACP-A; Flags: Precursor | 0.7829 | 17,124.2 | 27 | 19 | 1019 | 1,865,840.86 | 3.89 | Binding, Inhibits platelet aggregation, Antithrombotic and thrombolytic activities, Hydrolyzes the fibrinogen. |
SP|Q8JIV8.1|SL_DEIAC | Snaclec clone 2100755; AltName: Full = C-type lectin clone 2100755; Flags: Precursor | 0.671 | 17,944.1 | 25 | 25 | 464 | 1,649,481.55 | 3.44 | Binding, Modulation of platelet aggregation, or coagulation cascade. |
AAK26430.1 | Agkisasin-b [Deinagkistrodon acutus] | 0.7823 | 14,701.3 | 25 | 3 | 29 | 1,590,986.26 | 3.32 | Binding, Anticoagulant, Inhibits platelet aggregation, Antithrombotic and thrombolytic activities, Hydrolyzes the Aalpha-chain of fibrinogen. |
AFR11355.1 | Snake venom serine protease Da-36 [Deinagkistrodon acutus] | 0.5654 | 29,057.1 | 17 | 12 | 110 | 874,312.48 | 1.82 | Catalytic_activity, Blood coagulation factor activity, Serine elastase activity. |
AAN23124.1 | Agglucetin-alpha 1 subunit precursor [Deinagkistrodon acutus] | 0.8052 | 17,317.4 | 33 | 3 | 405 | 771,737.66 | 1.61 | Binding, Blood coagulation factor activity, Serine elastase activity. |
AAM22785.1 | Agkisacutacin B-chain [Deinagkistrodon acutus] | 0.8288 | 16,739.8 | 37 | 7 | 488 | 746,482.39 | 1.56 | Binding, Anticoagulant, Inhibits platelet aggregation, Antithrombotic and thrombolytic activities, Hydrolyzes the Aalpha-chain of fibrinogen. |
BAO23490.1 | Metalloproteinase [Deinagkistrodon acutus] | 0.4745 | 67,764.7 | 48 | 41 | 702 | 740,656.11 | 1.54 | Binding catalytic, Enzyme_regulator_activity, Metalloendoproteinase activity. |
SP|Q9IAX6.1|VM2M2_DEIAC | Zinc metalloproteinase/disintegrin | 0.6438 | 52,517.3 | 117 | 25 | 762 | 740,180.03 | 1.54 | Binding, enzyme_regulator_activity, inhibits Platelet aggregation, Cleaves fibrinogen, Cleaves collagen and VWF. |
PDB|3HDB|A | Chain A, Crystal Structure of Aahiv, A Metalloproteinase From Venom of Deinagkistrodon acutus | 0.8345 | 46,681.5 | 95 | 12 | 590 | 639,917.60 | 1.33 | Binding, Anticoagulant, Inhibits platelet aggregation, Antithrombotic and thrombolytic activities, Hydrolyzes the Aalpha-chain of fibrinogen. |
SP|Q9IAM1.2|SLUA_DEIAC | Snaclec agkisacutacin subunit A; Short = Agk-A; Flags: Precursor | 0.7895 | 17,109.1 | 21 | 3 | 287 | 502,397.74 | 1.05 | Binding, Anticoagulant, Inhibits platelet aggregation, Antithrombotic and thrombolytic activities, Hydrolyzes the Aalpha-chain of fibrinogen. |
ABB79955.1 | Agkihagin [Deinagkistrodon acutus] | 0.5789 | 67,572.2 | 50 | 48 | 969 | 500,625.222 | 1.045 | Binding, Enzyme_regulator_activity, Inhibits platelet aggregation. Hydrolyzes fibrinogen, Induces apoptosis and inhibits proliferation of endothelial cells. |
AAF76378.1 | Thrombin-like protein DAV-PA precursor [Deinagkistrodon acutus] | 0.6008 | 28,031.8 | 21 | 19 | 133 | 435,941.55 | 0.91 | Catalytic_activity, Fibrinogenolytic activities, Esterolysis and amidolytic activities. |
AEJ31983.1 | Phospholipase B [Crotalus adamanteus] | 0.5642 | 64,048.6 | 40 | 20 | 356 | 375,155.62 | 0.78 | Catalytic_activity, Hemolysis. |
AAL66391.1 | Antithrombin 1 A chain [Deinagkistrodon acutus] | 0.8608 | 17,950.1 | 51 | 19 | 445 | 332,370.033 | 0.69 | Platelet aggregation, Binds to human platelets, Promotes angiogenesis. |
JAG44660.1 | Glutaminyl-peptide cyclotransferase [Crotalus horridus] | 0.5815 | 42,369 | 16 | 16 | 116 | 320,694.98 | 0.67 | Binding |
AAM22789.1 | ACF 1/2 B-chain [Deinagkistrodon acutus] | 0.6712 | 16,924.9 | 26 | 5 | 229 | 265,179.49 | 0.55 | Binding, Anticoagulant, Inhibits platelet aggregation, Antithrombotic and thrombolytic activities, Hydrolyzes the Aalpha-chain of fibrinogen. |
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Huang, J.; Zhao, M.; Xue, C.; Liang, J.; Huang, F. Analysis of the Composition of Deinagkistrodon acutus Snake Venom Based on Proteomics, and Its Antithrombotic Activity and Toxicity Studies. Molecules 2022, 27, 2229. https://doi.org/10.3390/molecules27072229
Huang J, Zhao M, Xue C, Liang J, Huang F. Analysis of the Composition of Deinagkistrodon acutus Snake Venom Based on Proteomics, and Its Antithrombotic Activity and Toxicity Studies. Molecules. 2022; 27(7):2229. https://doi.org/10.3390/molecules27072229
Chicago/Turabian StyleHuang, Jin, Minrui Zhao, Chu Xue, Jiqiang Liang, and Fang Huang. 2022. "Analysis of the Composition of Deinagkistrodon acutus Snake Venom Based on Proteomics, and Its Antithrombotic Activity and Toxicity Studies" Molecules 27, no. 7: 2229. https://doi.org/10.3390/molecules27072229