Viperatoxin-II

a novel viper venom protein as an effective bactericidal agent

Ramar Perumal Samy, Bradley G. Stiles, Arunachalam Chinnathambi, M. E. Zayed, Sulaiman Ali Alharbi, Octavio Luiz Franco, Edward G. Rowan, Alan Prem Kumar, Lina H. K. Lim, Gautam Sethi

Research output: Contribution to journalArticle

8 Citations (Scopus)
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Abstract

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have become a rising threat to public health. There is an urgent need for development of promising new therapeutic agents against drug resistant bacteria like S. aureus. This report discusses purification and characterization of proteins from Indian Russell's viper snake venom. Novel 15-kDa proteins called "Viperatoxin" (VipTx-I and VipTx-II) were extracted from the whole venom and evaluated using in vitro antimicrobial experiments. The N-terminal amino acid sequence of "Viperatoxin" showed high sequence homology to daboiatoxin isolated from the same venom and also matched phospholipase A2 (PLA2) enzymes isolated from other snake venoms. In an in vitro plate assay, VipTx-II but not VipTx-I showed strong antimicrobial effects against S. aureus and Burkholderia pseudomallei (KHW & TES), Proteus vulgaris and P. mirabilis. The VipTx-II was further tested by a broth-dilution assay at 100-3.1μg/ml concentrations. The most potent bactericidal effect was found at the lowest dilutions (MICs of 6.25μg/ml) against B. pseudomallei, S. aureus and P. vulgaris (MICs of 12.25μg/ml). Electron microscopic investigation revealed that the protein-induced bactericidal potency was closely associated with pore formation and membrane damage, even at the lowest concentrations (<20μg/ml). The toxin caused a low level of cytotoxic effects as observed in human (THP-1) cells at higher concentrations. Molecular weight determinations of VipTx-II by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one major, along with a few minor bands. The results indicate that VipTx-II plays a significant role in bactericidal and membrane damaging effects in vitro. Non-cytotoxic properties on human cells highlight it as a promising candidate for further evaluation of antimicrobial potential in vivo.

Original languageEnglish
Pages (from-to)928-941
Number of pages14
JournalFEBS Open Bio
Volume5
Issue number1
DOIs
Publication statusPublished - 23 Oct 2015

Fingerprint

Viper Venoms
Burkholderia pseudomallei
Proteus vulgaris
Staphylococcus aureus
Snake Venoms
Venoms
Dilution
Assays
Russell's Viper
Mirabilis
Membranes
Proteins
Methicillin
Phospholipases A2
Public health
Methicillin-Resistant Staphylococcus aureus
Sequence Homology
Electrophoresis
Sodium Dodecyl Sulfate
Purification

Keywords

  • bactericidal
  • Daboia russelli russelli
  • Phospholipase A
  • Viperatoxin-I
  • Viperatoxin-II

Cite this

Samy, R. P., Stiles, B. G., Chinnathambi, A., Zayed, M. E., Alharbi, S. A., Franco, O. L., ... Sethi, G. (2015). Viperatoxin-II: a novel viper venom protein as an effective bactericidal agent. FEBS Open Bio, 5(1), 928-941. https://doi.org/10.1016/j.fob.2015.10.004
Samy, Ramar Perumal ; Stiles, Bradley G. ; Chinnathambi, Arunachalam ; Zayed, M. E. ; Alharbi, Sulaiman Ali ; Franco, Octavio Luiz ; Rowan, Edward G. ; Kumar, Alan Prem ; Lim, Lina H. K. ; Sethi, Gautam. / Viperatoxin-II : a novel viper venom protein as an effective bactericidal agent. In: FEBS Open Bio. 2015 ; Vol. 5, No. 1. pp. 928-941.
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Samy, RP, Stiles, BG, Chinnathambi, A, Zayed, ME, Alharbi, SA, Franco, OL, Rowan, EG, Kumar, AP, Lim, LHK & Sethi, G 2015, 'Viperatoxin-II: a novel viper venom protein as an effective bactericidal agent', FEBS Open Bio, vol. 5, no. 1, pp. 928-941. https://doi.org/10.1016/j.fob.2015.10.004

Viperatoxin-II : a novel viper venom protein as an effective bactericidal agent. / Samy, Ramar Perumal; Stiles, Bradley G.; Chinnathambi, Arunachalam; Zayed, M. E.; Alharbi, Sulaiman Ali; Franco, Octavio Luiz; Rowan, Edward G.; Kumar, Alan Prem; Lim, Lina H. K.; Sethi, Gautam.

In: FEBS Open Bio, Vol. 5, No. 1, 23.10.2015, p. 928-941.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Viperatoxin-II

T2 - a novel viper venom protein as an effective bactericidal agent

AU - Samy, Ramar Perumal

AU - Stiles, Bradley G.

AU - Chinnathambi, Arunachalam

AU - Zayed, M. E.

AU - Alharbi, Sulaiman Ali

AU - Franco, Octavio Luiz

AU - Rowan, Edward G.

AU - Kumar, Alan Prem

AU - Lim, Lina H. K.

AU - Sethi, Gautam

PY - 2015/10/23

Y1 - 2015/10/23

N2 - Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have become a rising threat to public health. There is an urgent need for development of promising new therapeutic agents against drug resistant bacteria like S. aureus. This report discusses purification and characterization of proteins from Indian Russell's viper snake venom. Novel 15-kDa proteins called "Viperatoxin" (VipTx-I and VipTx-II) were extracted from the whole venom and evaluated using in vitro antimicrobial experiments. The N-terminal amino acid sequence of "Viperatoxin" showed high sequence homology to daboiatoxin isolated from the same venom and also matched phospholipase A2 (PLA2) enzymes isolated from other snake venoms. In an in vitro plate assay, VipTx-II but not VipTx-I showed strong antimicrobial effects against S. aureus and Burkholderia pseudomallei (KHW & TES), Proteus vulgaris and P. mirabilis. The VipTx-II was further tested by a broth-dilution assay at 100-3.1μg/ml concentrations. The most potent bactericidal effect was found at the lowest dilutions (MICs of 6.25μg/ml) against B. pseudomallei, S. aureus and P. vulgaris (MICs of 12.25μg/ml). Electron microscopic investigation revealed that the protein-induced bactericidal potency was closely associated with pore formation and membrane damage, even at the lowest concentrations (<20μg/ml). The toxin caused a low level of cytotoxic effects as observed in human (THP-1) cells at higher concentrations. Molecular weight determinations of VipTx-II by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one major, along with a few minor bands. The results indicate that VipTx-II plays a significant role in bactericidal and membrane damaging effects in vitro. Non-cytotoxic properties on human cells highlight it as a promising candidate for further evaluation of antimicrobial potential in vivo.

AB - Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have become a rising threat to public health. There is an urgent need for development of promising new therapeutic agents against drug resistant bacteria like S. aureus. This report discusses purification and characterization of proteins from Indian Russell's viper snake venom. Novel 15-kDa proteins called "Viperatoxin" (VipTx-I and VipTx-II) were extracted from the whole venom and evaluated using in vitro antimicrobial experiments. The N-terminal amino acid sequence of "Viperatoxin" showed high sequence homology to daboiatoxin isolated from the same venom and also matched phospholipase A2 (PLA2) enzymes isolated from other snake venoms. In an in vitro plate assay, VipTx-II but not VipTx-I showed strong antimicrobial effects against S. aureus and Burkholderia pseudomallei (KHW & TES), Proteus vulgaris and P. mirabilis. The VipTx-II was further tested by a broth-dilution assay at 100-3.1μg/ml concentrations. The most potent bactericidal effect was found at the lowest dilutions (MICs of 6.25μg/ml) against B. pseudomallei, S. aureus and P. vulgaris (MICs of 12.25μg/ml). Electron microscopic investigation revealed that the protein-induced bactericidal potency was closely associated with pore formation and membrane damage, even at the lowest concentrations (<20μg/ml). The toxin caused a low level of cytotoxic effects as observed in human (THP-1) cells at higher concentrations. Molecular weight determinations of VipTx-II by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed one major, along with a few minor bands. The results indicate that VipTx-II plays a significant role in bactericidal and membrane damaging effects in vitro. Non-cytotoxic properties on human cells highlight it as a promising candidate for further evaluation of antimicrobial potential in vivo.

KW - bactericidal

KW - Daboia russelli russelli

KW - Phospholipase A

KW - Viperatoxin-I

KW - Viperatoxin-II

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DO - 10.1016/j.fob.2015.10.004

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SP - 928

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JO - FEBS Open Bio

JF - FEBS Open Bio

SN - 2211-5463

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Samy RP, Stiles BG, Chinnathambi A, Zayed ME, Alharbi SA, Franco OL et al. Viperatoxin-II: a novel viper venom protein as an effective bactericidal agent. FEBS Open Bio. 2015 Oct 23;5(1):928-941. https://doi.org/10.1016/j.fob.2015.10.004