ISSN : 1225-3480
동양달팽이의 EST 에서 serpin 유전자는 2가지 type이 추정되었다. Ns-serpin type 1 (partial) 서열의 코딩 영역은 총819 bp 이었으며, 272개의 아미노산으로 이루어져 있었으며, Ns-serpin type 2 (partial) 의 코딩영역은 총 555 bp, 185개의 아미노산으로 이루어져 있었다. Laminarin 처리 전에는Ns-serpin type 1 이 2개 발현되었으며 처리 후에는Ns-serpin type 2 가 1개 발현되고 있었다. BLAST 결과를바탕으로 유사도가 높은 17개의 참고 서열과 동양달팽이의Ns-serpin type 1, 2의 아미노산 서열을 MEGA6 프로그램의 clustalW 엔진을 이용하여 multiple sequence alignment 를 수행한 결과, 척색동물의 포유강 9종, 조류강 1 종, 양서강 1종과 척색동물인 해초강, 절지동물의 거미강 1종과 곤충강 1종이 같은 군으로 묶였으며, 연체동물문의 경우 동양달팽이의 두 가지 type 을 포함한 복족강 4종이 같은 군으로 묶이는 것을 확인하였다. Psipred 프로그램을 활용하여 예측한 2D 구조도 multiple align 및 phylodendrogram 결과와 연관이 있음을 확인할 수 있었다. EST를 통해 밝힌 동양달팽이의 두가지 type의 Ns-serpin 서열은 Aplysia californica 등의 근연종들의 서열과 유사하였다. 본 연구에서는 무척추동물에서의 선천성 면역과 연관이 있는 것으로 잘 알려진 두 종류의 serpin 유전자 서열을 동양달팽이 EST 결과에서 발굴 하였으며, 동양달팽이에서도 여러 가지 type의serpin 이 존재 할 수 있음을 입증하였다.
Serpins are a group of proteins involved in the regulation of serine and other type of proteases, and have been identified in many kinds of organisms from invertebrates to vertebrates. Serpins are known to regulate the proteolytic cascades of the innate immune pathways in addition to their roles in blood coagulation, angiogenesis, fibrinolysis, inflammation and tumor suppression. In this study, we have isolated two partial serpin gene fragments from expressed sequence tags (ESTs) of Nesiohelix samarangae. Dotplot analysis indicates that they are of two different types, Ns-serpin type 1 and Ns-serpin type 2. Ns-serpin type 1 has 819 bp coding region (272 amino acids), whereas Ns-serpin type 2 has 555 bp coding region (185 amino acids). Molecular phylogenetic analysis shows that the identified serpins have high similarities to their counterparts in the California see slug, Aplysia californica. Yet, the precise biological and immunological roles of these Ns-serpins remain to be further investigated using RNA interference and other molecular techniques.
Armstrong, P. B. (2001) "The contribution of proteinase inhibitors to immune defense." Trends Immunol., 22(1): 47-52.
Braun, R. C., K. T. Pedretti, T. L. Casavant, T. E. Scheetz, C. L. Birkett and C. A. Roberts (2001)"Parallelization of local BLAST service on workstation clusters." Future Generation Computer Systems, 17: 745-754.
Coyne, V. (2011) "The importance of ATP in the immune system of molluscs." Invertebrate Survival Journal, 8:48-55.
Cui, S., D. Zhang, S. Jiang, H. Pu, Y. Hu, H. Guo, M. Chen, T. Su and C. Zhu (2011) "A macrophage migration inhibitory factor like oxidoreductase from pearl oyster Pinctada fucata involved in innate immune responses." Fish Shellfish Immunol., 31(2):173-181.
Edgar, R. C. (2004) "MUSCLE: a multiple sequence alignment method with reduced time and space complexity." BMC Bioinformatics, 5: 113.
Edgar, R. C. (2004) "MUSCLE; multiple sequence alignment with high accuracy and high throughput."Nucleic Acids Res., 32: 1792-1797.
Gatto, M., L. Iaccarino, A. Ghirardello, N. Bassi, P. Pontisso, L. Punzi, Y. Shoenfeld and A. Doria (2013)"Serpins, immunity and autoimmunity: old molecules, new functions." Clin. Rev. Allergy Immunol., 45(2):267-280.
Jeffroy, F., F. Brulle and C. Paillard (2013) "Differential expression of genes involved in immunity and biomineralization during Brown Ring Disease development and shell repair in the Manila clam, Ruditapes philippinarum." J. Invertebr Pathol., 113(2): 129-136.
Jeong, J. E. and Y. S. Lee (2013) "Identification, sequence characterization and expression analysis of the arginine kinase gene in response to laminarin challenge from the Oriental land snail, Nesiohelix samarangae." Korean Journal of Malacology, 29(3):171-179.
Jones, D. T., W. R. Taylor and J. M. Thornton (1992)"The rapid generation of mutation data matrices from protein sequences." Comput Appl. Biosci., 8(3):275-282.
Kanost, M. R. (1999) "Serine proteinase inhibitors in arthropod immunity." Developmental and Comparative Immunology, 23: 291-301.
Maeda, M. and K. Nishizawa (1968) "Fine structure of laminaran of Eisenia bicyclis." J. Biochem., 63(2):199-206.
Maldonado-Aguayo, W., G. Nunez-Acuna, V. Valenzuela-Munoz, J. Chavez-Mardones and C. Gallardo-Escarate (2013) "Molecular characterization of two kazal-type serine proteinase inhibitor genes in the surf clam Mesodesma donacium exposed to Vibrio anguillarum." Fish Shellfish Immunol., 34(6):1448-1454.
Miura, Y., S. Kawabata, Y. Wakamiya, T. Nakamura and S. Iwanaga (1995) "A limulus intracellular coagulation inhibitor type 2. Purification, characterization, cDNA cloning, and tissue localization." J. Biol. Chem., 270(2): 558-565.
Ruby HP Law, Z. Qingwei and M. sheena (2006) "An overview of the serpin superfamily." Genome Biology, 7.
Sadaaki Iwanaga and B. L. Lee (2005) "Recent advances in the innate immunity of invertebrate animals."Journal of Biochemistry and Molecular Biology, 38(2): 128-150
Silverman, G. A., P. I. Bird, R. W. Carrell, F. C. Church, P. B. Coughlin, P. G. Gettins, J. A. Irving, D. A. Lomas, C. J. Luke, R. W. Moyer, P. A. Pemberton, E. Remold-O'Donnell, G. S. Salvesen, J. Travis and J. C. Whisstock (2001) "The serpins are an expanding superfamily of structurally similar but functionally diverse proteins. Evolution, mechanism of inhibition, novel functions, and a revised nomenclature." J. Biol. Chem., 276(36): 33293-33296.
Supajatura, V., H. Ushio, A. Nakao, S. Akira, K. Okumura, C. Ra and H. Ogawa (2002) "Differential responses of mast cell Toll-like receptors 2 and 4 in allergy and innate immunity." J. Clin. Invest., 109(10): 1351-1359.
Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei and S. Kumar (2011) "MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods." Mol. Biol. Evol., 28(10):2731-2739.
Tamura, K., G. Stecher, D. Peterson, A. Filipski and S. Kumar (2013) "MEGA6: Molecular Evolutionary Genetics Analysis version 6.0." Mol. Biol. Evol., 30(12): 2725-2729.
Tirloni, L., A. Seixas, A. Mulenga, S. Vaz Ida, Jr. and C. Termignoni (2014) "A family of serine protease inhibitors (serpins) in the cattle tick Rhipicephalus (Boophilus) microplus." Exp. Parasitol., 137: 25-34.
Wang, Z., J. Jian, Y. Lu, B. Wang and Z. Wu (2011) "A tandem-repeat galectin involved in innate immune response of the pearl oyster Pinctada fucata." Mar. Genomics, 4(3): 229-236.
Wei, X., J. Yang, J. Yang, X. Liu, M. Liu, D. Yang, J. Xu and X. Hu (2012) "A four-domain Kunitz-type proteinase inhibitor from Solen grandis is implicated in immune response." Fish Shellfish Immunol., 33(6):1276-1284.
Yu, Z., X. He, D. Fu and Y. Zhang (2011) "Two superoxide dismutase (SOD) with different subcellular localizations involved in innate immunity in Crassostrea hongkongensis." Fish Shellfish Immunol., 31(4): 533-539.
Zhang, H., L. Wang, L. Song, X. Song, B. Wang, C. Mu and Y. Zhang (2009) "A fibrinogen-related protein from bay scallop Argopecten irradians involved in innate immunity as pattern recognition receptor."Fish Shellfish Immunol., 26(1): 56-64.
