J Venom Res (2015), Vol 6, 11-18
Published online: 22 July 2015
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Enzymatic analysis of venom from Cuban scorpion Rhopalurus junceus
Alexis Díaz-Garcíaα*, Jenny Laura Ruiz-Fuentes ß, Arianna Yglesias-Riveraα, Hermis Rodríguez-Sánchez ß, Yanelis Riquenes Garloboα, Osmel Fleitas Martinez ß, José A Fraga Castroα
αResearch Department, Laboratories of Biofarmaceuticals and Chemistries Productions (LABIOFAM), Havana, Cuba
ßMicrobiology Department, Tropical Medicine Institute “Pedro Kouri”, Havana, Cuba
*Correspondence to: Alexis Díaz-García, Email: email@example.com, Tel.: +53 7 684 96 58, Fax: +53 7 683 03 26
Received: 05 February 2015; Revised: 22 May 2015; Accepted: 23 June 2015
© Copyright The Author(s). First Published by Library Publishing Media. This is an open access article, published under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0). This license permits non-commercial use, distribution and reproduction of the article, provided the original work is appropriately acknowledged with correct citation details.
Rhopalurus junceus scorpion venom has been identified as a natural extract with anticancer potential. Interestingly, this scorpion venom does not cause adverse symptoms in humans. However, there is scarce information about its composition and enzymatic activity. In this work, we determined the electrophoretic profile of the venom, the gelatinase and caseinolytic activity, and the phospholipase A2 (PLA2) and hemolytic activity. The effect of different venom doses (6.25, 12.5 and 25 mg/kg) on gastrocnemius muscle was also measured as CK and LDH activity in serum. The presence of hyaluronidase was determined by turbidimetric assay. The effect of different fractions obtained by gel filtration chromatography were evaluated at different concentrations (0.05, 0.1, 0.2, 0.4, 0.6mg/ml) against lung cancer cell A549 and lung normal cell MRC-5 using MTT assay. The electrophoretic profile demonstrated the presence of proteins bands around 67kDa, 43kDa, 18.4kDa and a majority band below 14.3kDa. The venom did not showed caseinolytic, gelatinase, PLA2 and hemolytic activity even at highest venom concentration used in the study. Scorpion venom only showed a significant toxic effect on gastrocnemius muscles identified by CK and LDH release after subcutaneous injection of 12.5 and 50mg/kg. Low molecular weight fractions (<4kDa) induced a significant cytotoxicity in A549 cells while high molecular weight proteins (45-60kDa) were responsible for hyaluronidase activity and toxic effect against MRC-5. Experiments indicate that Rhopalurus junceus scorpion venom has low enzymatic activity, which could contribute to the low toxic potential of this scorpion venom.
KEYWORDS: Rhopalurus junceus, scorpion venom, hyaluronidase, high molecular weight proteins, cancer cells