Proteomic Identification of Stonefish Synanceja verrucosa Venom

Tai-Yuan Chen¹, Yu-Huai Chang¹, Hsi-Pin Lin², Shui-Tein Chen³ and Deng-Fwu Hwang^{1, 4,}

¹Department of Food Science and Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, Taiwan, ROC

²Department of Food Science, Yuanpei University, Hsinchu, Taiwan, ROC

³Institute of Biological Chemistry and Genomic Research Center, Academia Sinica, Taipei, Taiwan, ROC

⁴Department of Health and Nutrition, Asia University, Taichung, Taiwan, ROC

Cite this paper:
Tai-Yuan Chen, Yu-Huai Chang, Hsi-Pin Lin, Shui-Tein Chen and Deng-Fwu Hwang. Proteomic Identification of Stonefish Synanceja verrucosa Venom. Journal of Food and Nutrition Research. 2015; 3(8):526-539. doi: 10.12691/jfnr-3-8-8

Abstract

Three venom toxins, neoverrucotoxin (neoVTX) α-subunit and β-subunit as well as verrucotoxin (VTX) β-subunit, were identified in the stonefish Synanceja verrucosa by SDS-PAGE, Native-PAGE and two-dimensional electrophoresis (2-DE) coupled with Matrix Assisted Laser Desorption Ionization-Quadrupole-Time-of-Flight (MALDI-Q-TOF). The venom estimated by Native-PAGE were 471, 358, 260 and 166 kDa. The predominate protein bands of crude venom were 84 and 75 kDa by SDS-PAGE. The crude venom protein fell in the region with pI values of 7-9 and molecular weights of 75-90 kDa by 2-DE. Peptide mass fingerprints (PMF) and MS/MS ions originated from MALDI-Q-TOF were used to identify the protein. Our results showed that the complete components of neoverrucotoxin (neoVTX) α-subunit and β-subunit as well as verrucotoxin (VTX) β-subunit were identified from SDS-PAGE and 2-DE patterns. Native-PAGE did not yield protein identifications but revealed the presence of protein complexes.

Keywords:
2-DE MALDI-Q-TOF neoverrucotoxin spine venom Synanceja verrucosa

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Church, J.E.and Hodgson, W.C. (2000). Dose-dependent cardiovascular effects of stonefish (Synanceja trachyrnis) venom. Toxicon, 38: 391-407.
[2]	Khoo, H.E., Yuen, R., Poh, C.H. and Tan, C.H. (1992). Biological activities of Synanceja horrida (stonefish) venom. Natural Toxins, 1: 54-60.
[3]	Ueda, A., Suzuki, M., Honma, T., Naga, H., Nagashima, Y. and Shiomi, K. (2006). Purification, properties and cDNA cloning of neoverrucotoxin (neoVTX), a hemolytic lethal factor from the stonefish Synanceja verrucosa venom. Biochimica et Biophysica Acta, 1760: 1713-1722.
[4]	Garnier, P., Goudey-Perriere, F., Breton, P., Dewulf, C., Petek, F. and Perriere, C. (1995). Enzymatic properties of the stonefish (Synanceja verrucosa, Bloch and Schneider, 1801) venom and purification of a lethal, hypotensive and cytolytic factor. Toxicon, 33: 143-155.
[5]	Garnier, P., Ducancel, F., Ogawa, T., Boulain, J.C., Goudey-Perriere, F., Perriere, C.and Menez, A. (1997). Complete amino-acid sequence of the beta-subunit of VTX from venom of the stonefish (Synanceja verrucosa) as identified from cDNA. Biochimica et Biophysica Acta, 1337: 1-5.
[6]	Breton, P., Delamancle, I., Buee, J., Goudey-Perriere, F. and Perriere C. (2002). Evidence for a neurotoxic activity in crude venom of stonefish (Synanceja verrucosa). Journal of Natural Toxins, 11: 305-318.
[7]	Madokoro, M., Ueda, A., Kiriake, A. and Shiomi, K. (2011). Properties and cDNA cloning of a hyaluronidase from the stonefish Synanceja verrucosa venom. Toxicon, 58: 285-292.
[8]	Garnier, P., Grosclaude, J.M., Goudy-Perriere, F., Gervat, V., Gayral, P., Jackquot, C. and Perriere, C. (1996). Presence of norepinephrine and their biogenic amines in stonefish venom. Journal of Chromatography B: Biomedical Sciences and Applications, 685: 364-369.
[9]	Khoo, H.E. (2002). Bioactive proteins from stonefish venom. Clinical and Experimental Pharmacology and Physiology, 29: 802-806.
[10]	Andrich, F., Carnielli, J.B., Cassoli, J.S., Lautner, R.Q., Santos, R.A.S., Pimenta, A.M.C., DeLima, M.F. and Figueiredos, S.S. (2010). A potent vasoactive cytolysin isolated from Scorpaema plumier scorpionfish venom. Toxicon, 56: 487-496.
[11]	Gomes, H.H., Menezes, T.N., Carnielli, J.B.T., Andrich, F., Evangelista, K.S., Chavez-Olortegui, C., Vassallo, D.Y. and Figueiredo, S.G. (2011). Stonefish antivenom neutralizes the inflammatory and cardiovascular effects induced by scorpionfish Scorpaena plumier venom. Toxicon, 57: 992-999.
[12]	Henzel, W.J., Billeci, T.M., Stults, J.T., Wong, S.C., Grimley, C. and Watanabe, C. (1993). Identifying proteins from two-dimensional gels by molecular mass searching of peptide fragments in protein sequence databases. Proceedings of National Academy of Sciences of the United States of America, 90: 5011-5015.
[13]	James, P., Quadroni, M., Carafoli, E. and Gonnet, G. (1993). Protein identification by mass profile fingerprinting. Biochemical and Biophysical Research Communications, 195: 58-64.
[14]	Mann, M., Højrup, P. and Roepstorff, P. (1993). Use of mass spectrometric molecular weight information to identify proteins in sequence databases. Biological Mass Spectrometry, 22: 338-345.
[15]	Pappin, D.J., Hojrup, P. and Bleasby, A.J. (1993). Rapid identification of proteins by peptide-mass fingerprinting. Current Biology, 3: 327-332.
[16]	Yates, J.R., Speicher, S., Griffin, P.R. and Hunkapiller, T. (1993). Peptide mass maps: a highly informative approach to protein identification. Analytical Biochemistry, 214: 397-408.
[17]	Tsai, Y.C. and Hwang, D.F. (1999). Recognition of sting fish. Monthly Fisheries Extention, 148: 25-29. (in Chinese)
[18]	Lin, S.P. and Hwang, D.F. (2007). Venom toxin in stingfish. Science Development, 420: 14-18. (in Chinese)
[19]	Guilhaus, M. (1995). Orthogonal acceleration time-of-flight mass spectrometry. Journal of Mass Spectrometry, 30: 1519-1532.
[20]	Yates, J.R. (1996). Protein structure analysis by mass spectrometry. Methods in Enzymology, 271: 351-377.
[21]	Ghadessy, F.J., Chen, D., Kini, R.M., Chung, M.C., Jeyaseelan, K., Khoo, H.E. and Yuen, R. (1996). Stonustoxin is a novel lethal factor from stonefish (Synanceja horrida) venom. Journal of Biological Chemistry, 271: 25575-25581.
[22]	Kiriake, A. and Shiomi, K. (2011). Some properties and cDNA cloning of proteinaceous toxins from two species of lionfish (Pterois antennata and Pterois lunulata). Toxicon, 58: 494-501.
[23]	Kiriake, A., Suzuki, Y., Nagashima, Y. and Shiomi, K. (2013). Proteinaceous toxins from three species of scorpaeniform fish (lionfish Pterois lunulata, devil stinger Inimicus japonicus and waspfish Hypodytes rubripinnis): Close similarity in properties and primary structures to stonefish toxins. Toxicon, 70: 184-193.
[24]	Chen, T.Y., Hsieh, Y.W., Tsai, Y.H., Shiau, C.Y. and Hwang, D.F. (2002). Identification of species and measurement of tetrodotoxin in dried dressed fillets of the puffer fish, Lagocephalus lunaris. Journal of Food Protection, 65: 1670-1673.
[25]	Tseng, H.W., Juan, H.F., Huang, H.C., Lin, J.Y., Sinchaikul, S., Lai, T.C., Chen, C.F., Chen, S.T. and Wang, G.J. (2006). Lipopolysaccharide-stimulated response in rat aortic endothelial cells by a systems biology approach. Proteomics, 6: 5915-5928.
[26]	Schelimer, R.P., Gillespie, E. and Lichtenstein, L.M. (1981). Release of histamine from human leukocytes stimulated with the tumor-promoting phorbol diesters. I. Characterization of response. Journal of Immunology, 126: 570-574.
[27]	Wittig, I., Braun, H.P. and Schägger, H. (2006). Blue native PAGE. Nature Protocols, 1: 418-428.
[28]	Wessels, H.J.C.T., Vogel, R.O., Heuvel, L.v.d., Smeitink, J.A., Rodenburg, R.J., Nijtmans, L.G. and Farhoud, M.H. (2009). LC-MS/MS as an alternative for SDS-PAGE in blue native analysis of protein complexes. Proteomics, 9: 4221-4228.
[29]	Shiomi, K., Hosaka, M. and Kikuchi, T. (1993). Properties of a lethal factor in stonefish Synanceia verrucosa venom. Nippon Suisan Gakkaishi, 59: 1099.
[30]	Poh, C.H., Yuen, R., Khoo, H.E., Chung, M., Gwee, M. and Gopalakrishnakone, P. (1991). Purification and partial characterization of stonustoxin (lethal factor) from Synanceja horrida venom. Comparative Biochemistry and Physiology-Part B: Biochemistry & Molecular Biology, 99: 793-798.
[31]	Austin, L., Gills, R.G. and Youatt, G. (1965). Stonefish venom: some biochemical and chemical observations. The Australian Journal of Experimental Biology and Medical Science, 43: 79-90.
[32]	Colasante, C., Meunier, F.A., Kreger, A.S. and Molgó, J. (1996). Selective depletion of clear synaptic vesicles and enhanced quantal transmitter release at frog motor nerve endings produced by trachynilysin, a protein toxin isolated from stonefish (Synanceia trachynis) venom. European Journal of Neuroscience, 8: 2149-2156.
[33]	Nagai, H. (2012). Marine protein toxins. In: Handbook of marine natural products, Vol. 1. (Eds) Fattorusso, E., Gerwick, W.H., Taglialatela-Scafati, O., Springer, New York, pp. 1389-1419.
[34]	Chen, D.S., Kini, R.M., Yuen, R. and Khoo, H.E. (1997). Haemolytic activity of stonustoxin from stonefish (Synanceja horrida) venom: Pore formation and the role of cationic amino acid residues. Biochemical Journal, 325: 685-691.
[35]	Khoo, H.E., Chen, D.S. and Yuen, R. (1998). The role of cationic amino acid residues in the lethal activity of stonustoxin from stonefish (Synanceja horrida) venom. Biochemistry and Molecular Biology International, 44: 643-646.
[36]	Khoo, H.E., Chen, D.S. and Yuen, R. (1998). Role of free thiol groups in the biological activities of stonustoxin, a lethal factor from stonefish (Synanceja horrida) venom. Toxicon, 36: 469-476.
[37]	Yew, W.S. and Khoo, H.E. (2000). The role of tryptophan residues in the hemolytic activity of stonustoxin, a lethal factor from stonefish (Synanceja horrida) venom. Biochimie, 82: 251-257.
[38]	Woo, J.S., Imm, J.H., Min, C.K., Kim, K.J., Cha, S.S. and Oh, B.H. (2006). Structural and functional insights into the B30.2/SPRY domain. The EMBO Journal, 25: 1353-1363.
[39]	Henry, J., Ribouchon, M.T., Offer, C. and Pontarotti, P. (1997). B30.2 like domain proteins: a growing family. Biochemical and Biophysical Research Communications, 235: 162-165.
[40]	Perfetto, L., Gheraadini, P.F., Davey, N.E., Diella, F., Helmer-Citterich, M. and Cesareni, G. (2013). Exploring the diversity of SPRY/B30.2-mediated interactions. Trends in Biochemical Sciences, 38: 38-46.