Abstract
Here, we report the purification and characterization of an acidic Asp49 phospholipase A2, named MVL-PLA2, with a molecular mass of 13,626.64 Da. The complete MVL-PLA2 cDNA was cloned from Macrovipera lebetina transmediterranea venom gland cDNA library. MVL-PLA2 possesses 122 amino acid residues, including 14 cysteines, and belongs to group II snake venom phospholipase A2 enzymes. MVL-PLA2 was not cytotoxic up to 2 muM and completely abolished cell adhesion and migration of various human tumor cells. Chemical modification with p-bromophenacyl bromide abolished the enzymatic activity of MVL-PLA2 without affecting its anti-tumor effect, suggesting the presence of 'pharmacological sites' distinct from the catalytic site in snake venom phospholipase A2. We demonstrated for the first time that the anti-tumor effect of MVL-PLA2 was mediated by alpha5beta1 and alphav-containing integrins. This finding may serve as starting point for structure-function relationship studies leading to design a new generation of specific anti-cancer drugs.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antigens, Neoplasm
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Antineoplastic Agents / chemistry
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Antineoplastic Agents / isolation & purification
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Antineoplastic Agents / pharmacology*
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Catalytic Domain / drug effects
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Cell Adhesion / drug effects
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Cell Line, Tumor / drug effects
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Cell Movement / drug effects
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Drug Screening Assays, Antitumor
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Fibrosarcoma / pathology
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Humans
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Integrin alpha5beta1 / antagonists & inhibitors
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Integrin alphaVbeta3 / antagonists & inhibitors
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Integrins / antagonists & inhibitors
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Melanoma / pathology
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Molecular Sequence Data
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Neoplasm Proteins / antagonists & inhibitors
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Phospholipases A2 / chemistry
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Phospholipases A2 / isolation & purification
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Phospholipases A2 / pharmacology*
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Sequence Alignment
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Sequence Homology, Amino Acid
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Structure-Activity Relationship
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Viper Venoms / chemistry
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Viper Venoms / enzymology*
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Viper Venoms / isolation & purification
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Viper Venoms / pharmacology
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Viperidae / metabolism*
Substances
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Antigens, Neoplasm
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Antineoplastic Agents
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Integrin alpha5beta1
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Integrin alphaVbeta3
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Integrins
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Neoplasm Proteins
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Viper Venoms
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integrin alphavbeta6
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MVL-PLA2, Macrovipera lebetina
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Phospholipases A2