Abstract
Phospholipase A2 (PLA2) is a major component in snake venoms and it is found in many different isoforms. To identify transcripts encoding different PLA2 isoforms, we developed a simple, rapid procedure. Total RNA was extracted from the venoms of three cottonmouth snakes and two diamondback rattlesnakes, and further reverse-transcribed into complementary DNA (cDNA). Using one pair of cottonmouth PLA2-specific primers and Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique, we identified 27 unique full-length PLA2 transcripts, including nine sequences identical to the previously documented ones in the database and one novel GIII-like PLA2. Two common transcripts respectively encoding Asp49 and Lys49 PLA2 isoforms were identified in all three cottonmouth venoms that contain more PLA2 transcripts than diamondback rattlesnake venoms. The placement of cloned PLA2 transcripts in snake venom PLA2s was further discussed by phylogenetic analysis. The procedure developed in this study paves the way for accelerated acquisition of transcriptome data on any other venom toxin families. The results obtained are crucial for insight into the structure and function of PLA2 isoforms for scientific and potential therapeutic purposes.
Highlights
Phospholipases (PLs) are ubiquitous enzymes present in all organisms and organs, and their primary function is to hydrolyze phospholipids
To investigate Phospholipase A2 (PLA2) transcripts, we selected a related group of cottonmouth snakes, and a related species from a different genus
These species were selected based on an abundance of PLA2 in their crude venoms, and the previous description of PLA2 transcriptome in venom glands and PLA2 toxins in crude venom [17,18,19]
Summary
Phospholipases (PLs) are ubiquitous enzymes present in all organisms and organs, and their primary function is to hydrolyze phospholipids. Depending on which ester linkage of phospholipid is hydrolyzed, PLs are classified into PLA1 , PLA2 , PLB, PLC and PLD [1]. Among these PLs, PLA2 s constitute a superfamily. Based on the primary structures and the organs to be expressed, PLA2 s are further divided into 16 groups [2,3]. Snake venom PLs are a secretory PLA2 and belong to at least group IA and II: IA (Elapidae), II (Viperidae). Group II venom PLA2 s are further divided into at least
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