Open Access Highly Accessed Research article

Quantitative high-throughput profiling of snake venom gland transcriptomes and proteomes (Ovophis okinavensis and Protobothrops flavoviridis)

Steven D Aird1*, Yutaka Watanabe1, Alejandro Villar-Briones1, Michael C Roy1, Kouki Terada2 and Alexander S Mikheyev1*

Author Affiliations

1 Okinawa Institute of Science and Technology, Tancha 1919-1, Onna-son, Kunigami-gun, Okinawa-ken 904-0412, Japan

2 Okinawa Prefectural Institute of Health and Environment, 2085 Ozato, Ozato Nanjo-shi, Okinawa-ken 901-1202, Japan

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BMC Genomics 2013, 14:790  doi:10.1186/1471-2164-14-790

Published: 14 November 2013

Abstract

Background

Advances in DNA sequencing and proteomics have facilitated quantitative comparisons of snake venom composition. Most studies have employed one approach or the other. Here, both Illumina cDNA sequencing and LC/MS were used to compare the transcriptomes and proteomes of two pit vipers, Protobothrops flavoviridis and Ovophis okinavensis, which differ greatly in their biology.

Results

Sequencing of venom gland cDNA produced 104,830 transcripts. The Protobothrops transcriptome contained transcripts for 103 venom-related proteins, while the Ovophis transcriptome contained 95. In both, transcript abundances spanned six orders of magnitude. Mass spectrometry identified peptides from 100% of transcripts that occurred at higher than contaminant (e.g. human keratin) levels, including a number of proteins never before sequenced from snakes. These transcriptomes reveal fundamentally different envenomation strategies. Adult Protobothrops venom promotes hemorrhage, hypotension, incoagulable blood, and prey digestion, consistent with mammalian predation. Ovophis venom composition is less readily interpreted, owing to insufficient pharmacological data for venom serine and metalloproteases, which comprise more than 97.3% of Ovophis transcripts, but only 38.0% of Protobothrops transcripts. Ovophis venom apparently represents a hybrid strategy optimized for frogs and small mammals.

Conclusions

This study illustrates the power of cDNA sequencing combined with MS profiling. The former quantifies transcript composition, allowing detection of novel proteins, but cannot indicate which proteins are actually secreted, as does MS. We show, for the first time, that transcript and peptide abundances are correlated. This means that MS can be used for quantitative, non-invasive venom profiling, which will be beneficial for studies of endangered species.

Keywords:
Transcriptome; Illumina; proteome; Mass spectrometry; Venom; Okinawa; Viperidae; Crotalinae; Toxins; Enzymes