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研究生:鍾文苑
研究生(外文):Wen-Yuan Chung
論文名稱:探討Nap2基因中含有RINGfingerdomain區域蛋白之功能
論文名稱(外文):Characterization og the function of RING domain of Nap2 gene
指導教授:簡靜香簡靜香引用關係
指導教授(外文):Chin-Hsiang Chien
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:Nap2區域蛋白之功能
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因為惡性腫瘤細胞表面的醣蛋白多會有sialylation程度增加的現象,因此有許多研究者想藉由唾液酸酶(sialidase)處理癌細胞,以降低癌細胞轉移以及入侵其他組織的能力。 在近年的研究中發現,大量表現唾液酸酶的細胞不單只會影響細胞表面醣蛋白唾液酸化的程度也會影響細胞內部的生理現象。
small nanH蛋白是梭狀芽胞桿菌的一種神經胺酸苷酶(neuraminidase),本實驗室以yeast two-hybrid篩選,發現了一個會與small nanH蛋白作用的未知功能基因,將它命名為Nap2(neuraminidase associated protein 2)。 經由北方墨點法分析(Northern blot)發現Nap2基因的轉錄產物全長應該是大於4kb。 我們在此研究的蛋白僅是NAP2蛋白中含有RING finger domain的部分氨基酸片段。 經過蛋白質序列比對後發現,R-NAP2蛋白的氨基酸序列包含了兩個可能的NLS序列和一個 RING finger domain。 共軛焦顯微鏡觀察發現EGFP-R-NAP2蛋白會分佈在細胞核外圍,且位在R-NAP2蛋白N端的NLS序列對R-NAP2分佈的影響較大,C端的NLS序列以及RING finger domain則較無顯著的影響。 以MTT assay分析發現,大量表現R-NAP2會抑制OVCAR-3、YM-1、HeLa和HEK293細胞株的細胞生長速率,但對HepG2/A2和Huh-7卻無顯著的影響。 根據西方墨點法(Western blot)和流式細胞儀的分析結果指出大量表現R-NAP2蛋白並不會誘導細胞走向細胞凋亡。 流式細胞儀(FACS)分析細胞週期的結果發現,大量表現R-NAP2 蛋白可能會造成細胞週期停頓(cell cycle arrest)在G2/M phase的現象。 由上述之實驗結果顯示,R-NAP2蛋白可能具有抑制腫瘤生長(tumor suppressor)的功能。
由於我們在此探討的都只是NAP2蛋白的部分氨基酸序列之功能,另外還帶有SPRY domain的NAP2蛋白全長是否也具有類似的功能或分佈在細胞的狀況還是值得我們進一步探討。
Because the cell surface of malignant cancer cells play a feature of hyper-sialylation of the glycoprotein, sialidase was thought to be a way to inhibit the metastasis and invasive ability of cancer cells. In recent years, it was found that sialidase may not only influence of the glycoproteins and glycolipids on the cell surface but also influence intra-cellular function.
Small nanH is a neuraminidase of Clostridium perfringens. After yeast two-hybrid screen, we found a novel gene which would interact with small nanH protein and it was named Nap2 (neuraminidase associated protein 2). The Northern blot analysis showed that the major full length transcript of Nap2 gene should be longer than 4kb. The domain we studied here was just part of the NAP2 protein sequence folded into RING finger domain, hence it was called RING domain of Nap2 (R-NAP2). There are two candidate NLS sequence and one RING finger domain in the protein sequence of R-NAP2. Confocal microscopy observation showed EGFP-R-NAP2 protein was localizated around nucleus. The NLS sequence near N-terminal but not the one near C-terminal, neither RING finger domain of R-NAP2, would affect the localization of R-NAP2 protein. In MTT assay analysis, transiently overexpression R-NAP2 protein would inhibit cell growth rate in OVCAR-3, YM-1, HeLa, and HEK 293 cell lines, but not in HepG2/A2 and Huh-7. Overexpression of R-NAP2 protein may not induce cell apoptosis according to the results of Western blot and FACS. On the other hand, the FACS analysis showed that overexpression of R-NAP2 may cause cell cycle arrest at G2/M phase. It showed that R-NAP2 protein may play a role as tumor suppressor.
But we still don’t know whether full length of NAP2 protein which contains another domain- SPRY domain would show similar function and localization as R-NAP2 protein dose.
Achyuthan, K. E., and Achyuthan, A. M. (2001). Comparative enzymology, biochemistry and pathophysiology of human exo-alpha-sialidases (neuraminidases). Comp Biochem Physiol B Biochem Mol Biol 129, 29-64.
Barlow, P. N., Luisi, B., Milner, A., Elliott, M., and Everett, R. (1994). Structure of the C3HC4 domain by 1H-nuclear magnetic resonance spectroscopy. A new structural class of zinc-finger. J Mol Biol 237, 201-211.
Bonten, E. J., Arts, W. F., Beck, M., Covanis, A., Donati, M. A., Parini, R., Zammarchi, E., and d'Azzo, A. (2000). Novel mutations in lysosomal neuraminidase identify functional domains and determine clinical severity in sialidosis. Hum Mol Genet 9, 2715-2725.
Cabezas, J.A., Reglero, A., and Calvo, P., (1983) Glycosidase (fucosides, galacosidase, glucosidase, hexoaminidases, and glucuronidase from some mollusks and vertebrates, and neuraminidase from virus). Int. J. Biochem. 15, 243-259.
Crocker, P.R., Kelm, S., Dubois, C., Martin, B., McWilliam, B.S., Shotten, K.M., Paulson, J.C., and Gordon, S., (1991) Purification and properties of sialoadhesion, a sialic acid-binding receptor of murine tissue macrophages. EMBO J. 10,1661-1669.
Chien, C. H., Huang, Y. C., and Chen, H. Y. (1997). Small neuraminidase gene of Clostridium perfringens ATCC 10543: cloning, nucleotide sequence, and production. Enzyme Microb Technol 20, 277-285.
Crennell, S. J., Garman, E. F., Laver, W. G., Vimr, E. R., and Taylor, G. L. (1993). Crystal structure of a bacterial sialidase (from Salmonella typhimurium LT2) shows the same fold as an influenza virus neuraminidase. Proc Natl Acad Sci U S A 90, 9852-9856.
Evans, D.G., Evans, D.J., Moulds, J.J., and Graham, D.U., (1988) N-acetylneuraminyllactose-binding fibrillar hemmagglutinin of Camplyovacter pylori: Aputative colonization factor antigen. Infect. Immun. 56, 2896-2906.
Freemont, P. S., Hanson, I. M., and Trowsdale, J. (1991). A novel cysteine-rich sequence motif. Cell 64, 483-484.
Freemont, P.S. (1993) The RING finger: a novel protein sequence motif related to the zinc finger. Ann NY Acad Sci. 684, 174-192
Freemont, P. S. (2000). RING for destruction? Curr Biol 10, R84-87.
Gaskell, A., Crennell, S., and Taylor, G. (1995). The three domains of a bacterial sialidase: a beta-propeller, an immunoglobulin module and a galactose-binding jelly-roll. Structure 3, 1197-1205.
Gillard, B.K., Thurmon, L.T., and Marcus, D.M. (1993) Variable subcellular localization of glycosphingolipids. Glycobiology 3, 57-67
Hirst, G.K., (1941) The agglutination of red cells by allanotic fluid of chick embryos infected with influenza virus. Sciences 94, 22-23
Kakugawa, Y., Wada, T., Yamaguchi, K., Yamanami, H., Ouchi, K., Sato, I., and Miyagi, T. (2002). Up-regulation of plasma membrane-associated ganglioside sialidase (Neu3) in human colon cancer and its involvement in apoptosis suppression. Proc Natl Acad Sci U S A 99, 10718-10723.
Kato, T., Wang, Y., Yamaguchi, K., Milner, C. M., Shineha, R., Satomi, S., and Miyagi, T. (2001). Overexpression of lysosomal-type sialidase leads to suppression of metastasis associated with reversion of malignant phenotype in murine B16 melanoma cells. Int J Cancer 92, 797-804.
Lovering, R., Hanson, I. M., Borden, K. L., Martin, S., O'Reilly, N. J., Evan, G. I., Rahman, D., Pappin, D. J., Trowsdale, J., and Freemont, P. S. (1993). Identification and preliminary characterization of a protein motif related to the zinc finger. Proc Natl Acad Sci U S A 90, 2112-2116.
Lowden, J. A., and O'Brien, J. S. (1979). Sialidosis: a review of human neuraminidase deficiency. Am J Hum Genet 31, 1-18.
Lukong, K. E., Elsliger, M. A., Chang, Y., Richard, C., Thomas, G., Carey, W., Tylki-Szymanska, A., Czartoryska, B., Buchholz, T., Criado, G. R., et al. (2000). Characterization of the sialidase molecular defects in sialidosis patients suggests the structural organization of the lysosomal multienzyme complex. Hum Mol Genet 9, 1075-1085.
Ponting, C., Schultz, J., and Bork, P. (1997) SPRY domains in ryanodine receptors (Ca2+-release channels). Trends Biochem. Sci. 22, 193-194.
Roggentin, P., Rothe, B., Kaper, J., Galen, J.,Lawrisuk, L., Vimr, E., and Schauer, R. (1989) Conserved sequences in bacterial and viral sislidases. Glycoconj. J. 6, 349-353.
Roggentin, T., Kleineidam, R. G., Schauer, R., and Roggentin, P. (1992). Effects of site-specific mutations on the enzymatic properties of a sialidase from Clostridium perfringens. Glycoconj J 9, 235-240.
Rothe, B., Roggentin, P., and Schauer, R. (1991). The sialidase gene from Clostridium septicum: cloning, sequencing, expression in Escherichia coli and identification of conserved sequences in sialidases and other proteins. Mol Gen Genet 226, 190-197.
Schauer, R. (1985) Sialic acids and their role as biological masks. Trends Biochem. Sci. 10, 357-360.
Schauer, R. (1982) Chemistry, metabolism, and biological functions of sialic acids. Adv Carbohydr Chem Biochem. 40, 131-234.
Schauer, R. (1992) Sialinsaurereuche schleime-bioaktive schmierrstoffe. Nachr. Chem. Tech. Lab. 40, 1227-1231.
Schauer, R., Kelm, S., Reuter, G., Roggentin, P., and Shaw, L. (1995) Biochemistry and role of sialic acids. In:Rosenberg, A. (Ed.), Biology of the Sialic Acids. Plenum Press, New York, 7-67.
Seyrantepe, V., Poupetova, H., Froissart, R., Zabot, M. T., Maire, I., and Pshezhetsky, A. V. (2003). Molecular pathology of NEU1 gene in sialidosis. Hum Mutat 22, 343-352.
Stamekovic, I., Sgroi, D., Aruffo, A.,SYM.S., and Anderson, T. (1991) The lymphocyte-B adhesion molecule Cd22 interacts with leukocyte common antigen CD45RO on T-cells andα-2,6-sialyltransferase, CD75, on B-cells. Cell 66, 1133-1144.
Stamekovic, I., Sgroi, D., and Aruffo, A. (1992) Cd22 binds to α-2,6- sialyltransferase-dependent epitopes in COS cells. Cell 68, 1003- 1004.
Suzuki, O., Nozawa, Y., and Abe, M. (2003). Sialic acids linked to glycoconjugates of Fas regulate the caspase-9-dependent and mitochondria-mediated pathway of Fas-induced apoptosis in Jurkat T cell lymphoma. Int J Oncol 23, 769-774.
Taylor, G. (1996). Sialidases: structures, biological significance and therapeutic potential. Curr Opin Struct Biol 6, 830-837.
Tokuyama, S., Moriya, S., Taniguchi, S., Yasui, A., Miyazaki, J., Orikasa, S., and Miyagi, T. (1997). Suppression of pulmonary metastasis in murine B16 melanoma cells by transfection of a sialidase cDNA. Int J Cancer 73, 410-415.
Tyers, M., and Jorgensen, P. (2000). Proteolysis and the cell cycle: with this RING I do thee destroy. Curr Opin Genet Dev 10, 54-64.
Wang, D., Li, Z., Messing, E. M., and Wu, G. (2002). Activation of Ras/Erk pathway by a novel MET-interacting protein RanBPM. J Biol Chem 277, 36216-36222.
Warner, T. G., Harris, R., McDowell, R., and Vimr, E. R. (1992). Photolabelling of Salmonella typhimurium LT2 sialidase. Identification of a peptide with a predicted structural similarity to the active sites of influenza-virus sialidases. Biochem J 285 ( Pt 3), 957-964.
Xie, Y., and Varshavsky, A. (1999). The E2-E3 interaction in the N-end rule pathway: the RING-H2 finger of E3 is required for the synthesis of multiubiquitin chain. Embo J 18, 6832-6844.
Yogeeswaran, G., and Salk, P. L. (1981). Metastatic potential is positively correlated with cell surface sialylation of cultured murine tumor cell lines. Science 212, 1514-1516.
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