- Proteomic characterization of two snake venoms: Naja naja atra and Agkistrodon halys

Proteomic characterization of two snake venoms: Naja naja atra and Agkistrodon halys
Shuting Li,* Jingqiang Wang,* Xumin Zhang,* Yan Ren,* Ning Wang,* Kang Zhao,* Xishu Chen,* Caifeng Zhao,* Xiaolei Li,* Jianmin Shao,* Jianning Yin,* Matthew B. West, Ningzhi Xu,* and Siqi Liu*1
*Beijing Genomics Institute (BGI), Chinese Academy of Sciences, I-Zone, Shunyi, Beijing 101300, China
†Beijing Proteomics Institute (BPI), I-Zone, Shunyi, Beijing 101300, China
‡The Department of Medicine, University of Louisville, Louisville, KY 40202, U.S.A.
1To whom correspondence should be addressed (email siqiliu/at/louisville.edu ).
Received March 4, 2004; Revised July 20, 2004; Accepted July 29, 2004.
Snake venom is a complex mixture of proteins and peptides, and a number of studies have described the biological properties of several venomous proteins. Nevertheless, a complete proteomic profile of venom from any of the many species of snake is not available. Proteomics now makes it possible to globally identify proteins from a complex mixture. To assess the venom proteomic profiles from Naja naja atra and Agkistrodon halys, snakes common to southern China, we used a combination strategy, which included the following four different approaches: (i) shotgun digestion plus HPLC with ion-trap tandem MS, (ii) one-dimensional SDS/PAGE plus HPLC with tandem MS, (iii) gel filtration plus HPLC with tandem MS and (iv) gel filtration and 2DE (two-dimensional gel electrophoresis) plus MALDI–TOF (matrix-assisted laser desorption ionization–time-of-flight) MS. In the present paper, we report the novel identification of 124 and 74 proteins and peptides in cobra and viper venom respectively. Functional analysis based upon toxin categories reveals that, as expected, cobra venom has a high abundance of cardio- and neurotoxins, whereas viper venom contains a significant amount of haemotoxins and metalloproteinases. Although approx. 80% of gel spots from 2DE displayed high-quality MALDI-TOF-MS spectra, only 50% of these spots were confirmed to be venom proteins, which is more than likely to be a result of incomplete protein databases. Interestingly, these data suggest that post-translational modification may be a significant characteristic of venomous proteins.
Keywords: Agkistrodon halys, Naja naja atra, proteome, snake, venom
Abbreviations: Agki, Agkistrodon halys, 1DE, one-dimensional gel electrophoresis (SDS/PAGE), 2DE, two-dimensional gel electrophoresis, DTT, dithiothreitol, 5′-END, 5′-ectonucleotidase, GF, gel filtration chromatography, HMK, haemorrhagic metalloproteinase kaouthiagin, LC-MS/MS, HPLC with tandem MS, MALDI–TOF MS, matrix-assisted laser-desorption ionization–time-of-flight, MM, molecular mass, Naja, Naja naja atra, TFA, trifluoroacetic acid
Biochem J. 2004 November 15; 384(Pt 1): 119–127. Copyright The Biochemical Society, London.

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