Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus)
Abstract
:1. Introduction
2. Results
2.1. Skull and Venom Gland Anatomical Comparisons
2.2. Proteomics
2.3. Enzymatic Assays
Fluorescent Determination of Matrix Metalloprotease and Kallikrein Activity
2.4. Fluorescent Determination of sPLA2 Activity
2.5. Procoagulation Analysis
3. Discussion
4. Materials and Methods
4.1. Venom Supplies
4.2. Micro-Computed Tomography (CT)
4.3. Magnetic Resonance Imaging
4.4. Proteomics
4.4.1. Nano HPLC-ESI-Triple Time of Flight (TOF) Mass Spectral Analysis
4.4.2. Protein Identification
4.4.3. Orbitap Elite Mass Spectrometer for SHOTGUNS
4.5. Enzymatic Activity Assays
4.5.1. Fluorescent Determination of Matrix Metalloprotease and Kallikrein Activity
4.5.2. Fluorescent Determination of PLA2 Activity
4.5.3. Enzymatic Statistical Analysis
4.6. Procoagulation Analysis
4.6.1. Whole Plasma Clotting
4.6.2. Antivenom Studies
4.6.3. Statistical Analysis
Whole Plasma Clotting EC50 Concentration and Asymptotic Time
Antivenom EC50 Concentration
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
CI | confidence interval |
aPTT | activated partial thromboplastin time |
SVMP | snake venom metalloprotease |
SDS-PAGE | sodium dodecyl sulfate polyacrylamide gel electrophoresis |
CRiSP | cysteine rich secretory protein |
HPLC | high pressure liquid chromatography |
SD | standard deviations |
ACN | acetonitrile |
1D | one dimensional |
2D | two dimensional |
3FTx | three finger toxin |
PLA2 | phospholipase A2 |
References
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Concentration | Without Antivenom | With Antivenom | ||||||
---|---|---|---|---|---|---|---|---|
D. typus | T. mossambicanus | D. typus | T. mossambicanus | |||||
Mean | SD | Mean | SD | Mean | SD | Mean | SD | |
20 µg/mL | 11.27 | 0.06 | 7.57 | 0.15 | 20.73 | 1.07 | 9.6 | 0.17 |
10 µg/mL | 15.95 | 1.88 | 9.55 | 0.17 | 155 | 5.2 | 16 | 0.17 |
6.66 µg/mL | 19.34 | 0.21 | 11.20 | 0.34 | 214.56 | 21.28 | 21.16 | 0.23 |
5 µg/mL | 21.22 | 0.22 | 12.43 | 0.08 | 255 | 10.56 | 35.13 | 1.32 |
3.33 µg/mL | 24.72 | 0.22 | 14.95 | 0.65 | 343.2 | 8.97 | 43.33 | 1.86 |
2.5 µg/mL | 27.63 | 0.14 | 16.45 | 0.18 | 432.3 | 35.32 | 53.43 | 0.11 |
2 µg/mL | 30.91 | 0.47 | 18.27 | 0.13 | 444.93 | 23.43 | 58.46 | 1.29 |
1.33 µg/mL | 36.32 | 1.61 | 20.50 | 0.58 | 525.86 | 51.19 | 75.43 | 2.15 |
1 µg/mL | 39.60 | 0.54 | 24.30 | 0.74 | 594.86 | 61.15 | 83.33 | 3.86 |
0.66 µg/mL | 43.91 | 0.56 | 27.44 | 1.03 | 735.3 | 6.94 | 100.8 | 1.97 |
0.4 µg/mL | 53.77 | 6.21 | 31.67 | 0.19 | 790.36 | 9.41 | 128.4 | 0.52 |
0.25 µg/mL | 61.95 | 7.95 | 35.91 | 0.64 | 966.76 | 35.88 | 163.23 | 2.6 |
0.125 µg/mL | 83.31 | 6.51 | 54.89 | 0.14 | 999 | 0 | 226.33 | 2.6 |
0.05 µg/mL | 146.65 | 5.23 | 80.44 | 0.52 | 999 | 0 | 381.23 | 4.71 |
Species | Coefficient Estimates | ||
---|---|---|---|
Aym ϕ1 | R0 ϕ2 | lrc ϕ3 | |
D. typus | 9.095161 (s) | 37.83835 (s) | −0.6711807 |
T. mossambicanus | 2.068652 (s) | 23.17609 (s) | −0.8289048 |
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Debono, J.; Dobson, J.; Casewell, N.R.; Romilio, A.; Li, B.; Kurniawan, N.; Mardon, K.; Weisbecker, V.; Nouwens, A.; Kwok, H.F.; et al. Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus). Toxins 2017, 9, 171. https://doi.org/10.3390/toxins9050171
Debono J, Dobson J, Casewell NR, Romilio A, Li B, Kurniawan N, Mardon K, Weisbecker V, Nouwens A, Kwok HF, et al. Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus). Toxins. 2017; 9(5):171. https://doi.org/10.3390/toxins9050171
Chicago/Turabian StyleDebono, Jordan, James Dobson, Nicholas R. Casewell, Anthony Romilio, Bin Li, Nyoman Kurniawan, Karine Mardon, Vera Weisbecker, Amanda Nouwens, Hang Fai Kwok, and et al. 2017. "Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus)" Toxins 9, no. 5: 171. https://doi.org/10.3390/toxins9050171
APA StyleDebono, J., Dobson, J., Casewell, N. R., Romilio, A., Li, B., Kurniawan, N., Mardon, K., Weisbecker, V., Nouwens, A., Kwok, H. F., & Fry, B. G. (2017). Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus). Toxins, 9(5), 171. https://doi.org/10.3390/toxins9050171