Crovirin, a Snake Venom Cysteine-Rich Secretory Protein (CRISP) with Promising Activity against Trypanosomes and Leishmania

Abstract

BackgroundThe neglected human diseases caused by trypanosomatids are currently treated with toxic therapy with limited efficacy. In search for novel anti-trypanosomatid agents, we showed previously that the Crotalus viridis viridis (Cvv) snake venom was active against infective forms of Trypanosoma cruzi. Here, we describe the purification of crovirin, a cysteine-rich secretory protein (CRISP) from Cvv venom with promising activity against trypanosomes and Leishmania.Methodology/Principal FindingsCrude venom extract was loaded onto a reverse phase analytical (C8) column using a high performance liquid chromatographer. A linear gradient of water/acetonitrile with 0.1% trifluoroacetic acid was used. The peak containing the isolated protein (confirmed by SDS-PAGE and mass spectrometry) was collected and its protein content was measured. T. cruzi trypomastigotes and amastigotes, L. amazonensis promastigotes and amastigotes and T. brucei rhodesiense procyclic and bloodstream trypomastigotes were challenged with crovirin, whose toxicity was tested against LLC-MK2 cells, peritoneal macrophages and isolated murine extensor digitorum longus muscle. We purified a single protein from Cvv venom corresponding, according to Nano-LC MS/MS sequencing, to a CRISP of 24,893.64 Da, henceforth referred to as crovirin. Human infective trypanosomatid forms, including intracellular amastigotes, were sensitive to crovirin, with low IC50 or LD50 values (1.10–2.38 µg/ml). A considerably higher concentration (20 µg/ml) of crovirin was required to elicit only limited toxicity on mammalian cells.ConclusionsThis is the first report of CRISP anti-protozoal activity, and suggests that other members of this family might have potential as drugs or drug leads for the development of novel agents against trypanosomatid-borne neglected diseases.

Recent updates and perspectives on leishmaniasis

“…Crovirin, a cysteinerich secretory protein (CRISP), extracted from Crotalus viridis viridis snake venom, used with a highperformance liquid chromatographer, had promising results against trypanosomes and leishmania [54]. This family might have potential as drugs or drug leads for the development of novel agents against these protozoa.…”

Cysteine‐rich buccal gland protein suppressed the proliferation, migration and invasion of hela cells through akt pathway

“…To date, CRISPs have been found in the venom glands of snakes , salivary glands of lizards and reproductive tracts of mammals . In the mammals, CRISPs are proved to be associated with sperm–egg fusion during the fertilization process ; while in the reptiles, CRISPs are reported to act as ion channel blockers .…”

“…Among these molecules, lots of studies have shown that disintegrins isolated from snakes are capable of inhibiting the proliferation, adhesion and migration of tumor cells effectively . Besides disintegrins, the cysteine‐rich secretory proteins (CRISPs) are also one of the most important constituents present in the venom glands of snakes . Usually, CRISPs from snakes are characterized by 16 conserved cysteines which locate at the carboxyl terminal of the proteins .…”

“…Different with disintegrins, most CRISPs in the venom glands of snakes are able to block various channels including voltage‐gated calcium channels, potassium channels, cyclic nucleotide‐gated ion channels and ryanodine receptors . Furthermore, CRISPs are also reported to cause myotoxicity in murine gastrocnemius muscle, to induce immune responses as well as to inhibit the infective forms of trypanosomatid parasites . Very recently, a CRSIP named as ES‐CRISP purified from the venom of Echis carinatus sochureki exhibited anti‐angiogenic activities by direct interaction with endothelial cells .…”

Isolation and characterization of an anti‐leishmanial disintegrin from Cerastes cerastes venom

“…The antiparasitic and antibacterial activities of several venoms were mainly ascribed to PLA 2 and l ‐amino acid oxidase ( l ‐AAO) . Nevertheless, Crovirin, a Snake Venom Cysteine‐Rich Secretory Protein (CRISP), was characterized with an anti‐protozoal activity against trypanosomes and Leishmania suggesting the anti‐parasitic potential of non‐enzymatic venom compounds …”