Abstract
Centruroides tecomanus is a Mexican scorpion endemic of the State of Colima, that causes human fatalities. This communication describes a proteome analysis obtained from milked venom and a transcriptome analysis from a cDNA library constructed from two pairs of venom glands of this scorpion. High perfomance liquid chromatography separation of soluble venom produced 80 fractions, from which at least 104 individual components were identified by mass spectrometry analysis, showing to contain molecular masses from 259 to 44,392 Da. Most of these components are within the expected molecular masses for Na+- and K+-channel specific toxic peptides, supporting the clinical findings of intoxication, when humans are stung by this scorpion. From the cDNA library 162 clones were randomly chosen, from which 130 sequences of good quality were identified and were clustered in 28 contigs containing, each, two or more expressed sequence tags (EST) and 49 singlets with only one EST. Deduced amino acid sequence analysis from 53% of the total ESTs showed that 81% (24 sequences) are similar to known toxic peptides that affect Na+-channel activity, and 19% (7 unique sequences) are similar to K+-channel especific toxins. Out of the 31 sequences, at least 8 peptides were confirmed by direct Edman degradation, using components isolated directly from the venom. The remaining 19%, 4%, 4%, 15% and 5% of the ESTs correspond respectively to proteins involved in cellular processes, antimicrobial peptides, venom components, proteins without defined function and sequences without similarity in databases. Among the cloned genes are those similar to metalloproteinases.
Highlights
Scorpionism is the epidemiologic word commonly used to define human accidents caused by scorpion stings, which occurs in many parts of the world, but in seven different geographical regions: three in Africa (North, Sahelian region and South), Asia Minor, South of India, Mexico and South America, where circa 2.3 billions of human persons are at risk [1].There are approximately 1,500 different species of scorpion described in the world, from which over 200 are found in Mexico [2]
The results of cDNA analysis shows that the venom glands of scorpions have many components related to cellular processes, protein synthesis, protein trafficking and others [31,32,33,34,35,36,37,38]. In this communication we report results of a transcriptome analysis obtained from a cDNA library prepared from the venom glands of two C. tecomanus scorpions, as well as a proteomic analysis from the soluble venom of this species
The molecular mass distribution of the venom components found in the 80 fractions are reported, clustered within different intervals of molecular weights, mostly 1000 Da apart from each other
Summary
Scorpionism is the epidemiologic word commonly used to define human accidents caused by scorpion stings, which occurs in many parts of the world, but in seven different geographical regions: three in Africa (North, Sahelian region and South), Asia Minor, South of India, Mexico and South America, where circa 2.3 billions of human persons are at risk [1].There are approximately 1,500 different species of scorpion described in the world, from which over 200 are found in Mexico [2]. Earlier taxonomic classification [4] of this scorpion placed this species within the family Buthidae, genus Centruroides, species limpidus, sub-especies tecomanus. Under this denomination (Centruroides limpidus tecomanus), a few articles related to venom components [6,7,8] were published by our group, as it will be shortly described below. Ponce-Saavedra et al [9] renamed it, as a bona fide species (Centruroides tecomanus), here thereafter abbreviated C. tecomanus It plays an important health problem in the country, especially in the State of Colima [10]
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