簡易檢索 / 詳目顯示

研究生: 邱柏強
Bo-Chiang Chiu
論文名稱: 脂聯素暨第一型脂聯素受體訊息傳遞之研究
The Study of Signaling Transduction in Adiponectin/AdipoR1 Pathway
指導教授: 林炎壽
Lin, Yenshou
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2012
畢業學年度: 100
語文別: 英文
論文頁數: 48
中文關鍵詞: 脂聯素代謝症候群第一型脂聯素受體過氧化體增殖劑活化受器穿膜蛋白43
英文關鍵詞: adiponectin, metabolic syndromes, AdipoR1, PPARs, TMEM43
論文種類: 學術論文
相關次數: 點閱:31下載:3
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • 脂聯素(adiponectin)是一種由脂肪組織所分泌的細胞激素(adipokines),由於它能促進個體對胰島素的敏感度,對於第二型糖尿病、冠狀動脈疾病以及高血壓等代謝症候群具有顯著得改善效果,因此在近幾年吸引了許多人投入研究。然而脂聯素的訊息傳遞路徑仍充滿許多未知,因此在本研究當中我們試圖找出脂聯素的訊息傳遞路徑中能與第一型脂聯素受體(AdipoR1)交互作用的傳導蛋白。我們採用了酵母雙雜交(yeast two hybrid)以及免疫沉澱(immunoprecipitation)兩種方法,其中從免疫沉澱實驗中的質譜儀定序結果中,我們找到穿膜蛋白43(TMEM 43)。我們發現在螢光顯微鏡下可觀察到細胞中標記紅螢光蛋白的重組TMEM43(RFP-TMEM43)與標記綠螢光蛋白的重組AdipoR1(GFP-AdipoR1)的分布似乎有共定位的現象。進一步共免疫沉澱的實驗加以驗證也顯示了標記FLAG的重組TMEM43 (FLAG-TMEM43) 與標記綠螢光蛋白的重組AdipoR1(GFP-AdipoR1) 之間有顯著的交互作用。另外在功能方面,因為之前有報告發現TMEM43的表現會受到伽瑪型過氧化體增殖劑活化受器(PPAR-γ)所活化。而過氧化體增殖劑活化受器已被確定和脂聯素的訊息傳遞路徑有關,因此我們假設TMEM43可能透過過氧化體增殖劑活化受器的調節而涉及脂聯素的訊息傳遞路徑中。期待本研究能釐清脂聯素的訊息傳遞路徑,以利於代謝症候群治療方法的未來發展。

    Adiponectin which is one of adipokines has attracted much attention in these years because of its significant effects to improve metabolic syndromes, such as type 2 diabetes, coronary artery disease, and hypertension. Nevertheless, the signal transduction pathway of adiponectin remains largely unknown. Utilizing two approaches, yeast two hybrid and immunoprecipitation, we began to explore the AdipoR1 binding partners which might be involved in adiponectin/AdipoR1 signaling pathway. TMEM43 was found in the immunoprecipitation recovery database of MS-SPEC results. Expression of RFP-TMEM43 and GFP-AdipoR1 in many cell types seems to colocalize under fluorescent microscope. Coimmunoprecipitation experiments were further employed to demonstrate that FLAG-TMEM43 significantly binds to GFP-AdipoR1. Previous studies considered that TMEM43 might be activated by PPAR-γ which involved in regulating cellular lipid metabolism in adiponectin/AdipoR1 signaling pathway. It is plausible that TMEM43 could play an important role in adiponectin/AdipoR1/PPARs signaling pathway. Hopefully, this study could provide new therapeutic target(s) for the drugs development to ameliorate metabolism related diseases.

    Abstract (Chinese) 1 Abstract (English) 3 Introduction 4 Materials and methods 9 Results 16 Discussion 21 References 24 Figures 31 Appendix 47

    Arita Y, Kihara S, Ouchi N, Takahashi M, Maeda K, Miyagawa J, Hotta K, Shimomura I, Nakamura T, Miyaoka K, Kuriyama H, Nishida M, Yamashita S, Okubo K, Matsubara K, Muraguchi M, Ohmoto Y, Funahashi T, Matsuzawa Y (1999) Paradoxical decrease of an adipose-specific protein, adiponectin, in obesity. Biochem Biophys Res Commun 257:79-83.
    Barger PM, Browning AC, Garner AN, Kelly DP (2001) p38 mitogen-activated protein kinase activates peroxisome proliferator-activated receptor alpha: a potential role in the cardiac metabolic stress response. J Biol Chem 276:44495-44501.
    Bateman A, Coin L, Durbin R, Finn RD, Hollich V, Griffiths-Jones S, Khanna A, Marshall M, Moxon S, Sonnhammer EL, Studholme DJ, Yeats C, Eddy SR (2004) The Pfam protein families database. Nucleic Acids Res 32:D138-141.
    Bengtsson L, Otto H (2008) LUMA interacts with emerin and influences its distribution at the inner nuclear membrane. J Cell Sci 121:536-548.
    Berg AH, Combs TP, Du X, Brownlee M, Scherer PE (2001) The adipocyte-secreted protein Acrp30 enhances hepatic insulin action. Nat Med 7:947-953.
    Berg AH, Combs TP, Scherer PE (2002) ACRP30/adiponectin: an adipokine regulating glucose and lipid metabolism. Trends Endocrinol Metab 13:84-89.
    Dreger M, Bengtsson L, Schoneberg T, Otto H, Hucho F (2001) Nuclear envelope proteomics: novel integral membrane proteins of the inner nuclear membrane. Proc Natl Acad Sci U S A 98:11943-11948.
    Flier JS (2004) Obesity wars: molecular progress confronts an expanding epidemic. Cell 116:337-350.
    Friedman JM (2000) Obesity in the new millennium. Nature 404:632-634.
    Fruebis J, Tsao TS, Javorschi S, Ebbets-Reed D, Erickson MR, Yen FT, Bihain BE, Lodish HF (2001) Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice. Proc Natl Acad Sci U S A 98:2005-2010.
    Heald M, Cawthorne MA (2011) Dual acting and pan-PPAR activators as potential anti-diabetic therapies. Handb Exp Pharmacol 35-51.
    Hotamisligil GS (1999) The role of TNFalpha and TNF receptors in obesity and insulin resistance. J Intern Med 245:621-625.
    Hu E, Liang P, Spiegelman BM (1996) AdipoQ is a novel adipose-specific gene dysregulated in obesity. J Biol Chem 271:10697-10703.
    Kadowaki T, Yamauchi T, Kubota N, Hara K, Ueki K, Tobe K (2006) Adiponectin and adiponectin receptors in insulin resistance, diabetes, and the metabolic syndrome. J Clin Invest 116:1784-1792.
    Lemay DG, Hwang DH (2006) Genome-wide identification of peroxisome proliferator response elements using integrated computational genomics. J Lipid Res 47:1583-1587.
    Liang WC, Mitsuhashi H, Keduka E, Nonaka I, Noguchi S, Nishino I, Hayashi YK (2011) TMEM43 mutations in Emery-Dreifuss muscular dystrophy-related myopathy. Ann Neurol 69:1005-1013.
    Long X, Lin Y, Ortiz-Vega S, Yonezawa K, Avruch J (2005) Rheb binds and regulates the mTOR kinase. Curr Biol 15:702-713.
    Maeda K, Okubo K, Shimomura I, Funahashi T, Matsuzawa Y, Matsubara K (1996) cDNA cloning and expression of a novel adipose specific collagen-like factor, apM1 (AdiPose Most abundant Gene transcript 1). Biochem Biophys Res Commun 221:286-289.
    Maeda N, Takahashi M, Funahashi T, Kihara S, Nishizawa H, Kishida K, Nagaretani H, Matsuda M, Komuro R, Ouchi N, Kuriyama H, Hotta K, Nakamura T, Shimomura I, Matsuzawa Y (2001) PPARgamma ligands increase expression and plasma concentrations of adiponectin, an adipose-derived protein. Diabetes 50:2094-2099.
    Mao X, Kikani CK, Riojas RA, Langlais P, Wang L, Ramos FJ, Fang Q, Christ-Roberts CY, Hong JY, Kim RY, Liu F, Dong LQ (2006) APPL1 binds to adiponectin receptors and mediates adiponectin signalling and function. Nat Cell Biol 8:516-523.
    Merner ND, Hodgkinson KA, Haywood AF, Connors S, French VM, Drenckhahn JD, Kupprion C, Ramadanova K, Thierfelder L, McKenna W, Gallagher B, Morris-Larkin L, Bassett AS, Parfrey PS, Young TL (2008) Arrhythmogenic right ventricular cardiomyopathy type 5 is a fully penetrant, lethal arrhythmic disorder caused by a missense mutation in the TMEM43 gene. Am J Hum Genet 82:809-821.
    Michael LF, Wu Z, Cheatham RB, Puigserver P, Adelmant G, Lehman JJ, Kelly DP, Spiegelman BM (2001) Restoration of insulin-sensitive glucose transporter (GLUT4) gene expression in muscle cells by the transcriptional coactivator PGC-1. Proc Natl Acad Sci U S A 98:3820-3825.
    Nakano Y, Tobe T, Choi-Miura NH, Mazda T, Tomita M (1996) Isolation and characterization of GBP28, a novel gelatin-binding protein purified from human plasma. J Biochem 120:803-812.
    Pajvani UB, Du X, Combs TP, Berg AH, Rajala MW, Schulthess T, Engel J, Brownlee M, Scherer PE (2003) Structure-function studies of the adipocyte-secreted hormone Acrp30/adiponectin. Implications fpr metabolic regulation and bioactivity. J Biol Chem 278:9073-9085.
    Puigserver P, Rhee J, Lin J, Wu Z, Yoon JC, Zhang CY, Krauss S, Mootha VK, Lowell BB, Spiegelman BM (2001) Cytokine stimulation of energy expenditure through p38 MAP kinase activation of PPARgamma coactivator-1. Mol Cell 8:971-982.
    Scherer PE, Williams S, Fogliano M, Baldini G, Lodish HF (1995) A novel serum protein similar to C1q, produced exclusively in adipocytes. J Biol Chem 270:26746-26749.
    Shapiro L, Scherer PE (1998) The crystal structure of a complement-1q family protein suggests an evolutionary link to tumor necrosis factor. Curr Biol 8:335-338.
    Shimomura I, Funahashi T, Takahashi M, Maeda K, Kotani K, Nakamura T, Yamashita S, Miura M, Fukuda Y, Takemura K, Tokunaga K, Matsuzawa Y (1996) Enhanced expression of PAI-1 in visceral fat: possible contributor to vascular disease in obesity. Nat Med 2:800-803.
    Shulman GI (2000) Cellular mechanisms of insulin resistance. J Clin Invest 106:171-176.
    Steppan CM, Bailey ST, Bhat S, Brown EJ, Banerjee RR, Wright CM, Patel HR, Ahima RS, Lazar MA (2001) The hormone resistin links obesity to diabetes. Nature 409:307-312.
    Tomas E, Tsao TS, Saha AK, Murrey HE, Zhang Cc C, Itani SI, Lodish HF, Ruderman NB (2002) Enhanced muscle fat oxidation and glucose transport by ACRP30 globular domain: acetyl-CoA carboxylase inhibition and AMP-activated protein kinase activation. Proc Natl Acad Sci U S A 99:16309-16313.
    Tsao TS, Lodish HF, Fruebis J (2002) ACRP30, a new hormone controlling fat and glucose metabolism. Eur J Pharmacol 440:213-221.
    Waki H, Yamauchi T, Kamon J, Ito Y, Uchida S, Kita S, Hara K, Hada Y, Vasseur F, Froguel P, Kimura S, Nagai R, Kadowaki T (2003) Impaired multimerization of human adiponectin mutants associated with diabetes. Molecular structure and multimer formation of adiponectin. J Biol Chem 278:40352-40363.
    White RT, Damm D, Hancock N, Rosen BS, Lowell BB, Usher P, Flier JS, Spiegelman BM (1992) Human adipsin is identical to complement factor D and is expressed at high levels in adipose tissue. J Biol Chem 267:9210-9213.
    Yamauchi T, Kamon J, Ito Y, Tsuchida A, Yokomizo T, Kita S, Sugiyama T, Miyagishi M, Hara K, Tsunoda M, Murakami K, Ohteki T, Uchida S, Takekawa S, Waki H, Tsuno NH, Shibata Y, Terauchi Y, Froguel P, Tobe K, Koyasu S, Taira K, Kitamura T, Shimizu T, Nagai R, Kadowaki T (2003a) Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature 423:762-769.
    Yamauchi T, Kamon J, Minokoshi Y, Ito Y, Waki H, Uchida S, Yamashita S, Noda M, Kita S, Ueki K, Eto K, Akanuma Y, Froguel P, Foufelle F, Ferre P, Carling D, Kimura S, Nagai R, Kahn BB, Kadowaki T (2002) Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase. Nat Med 8:1288-1295.
    Yamauchi T, Kamon J, Waki H, Imai Y, Shimozawa N, Hioki K, Uchida S, Ito Y, Takakuwa K, Matsui J, Takata M, Eto K, Terauchi Y, Komeda K, Tsunoda M, Murakami K, Ohnishi Y, Naitoh T, Yamamura K, Ueyama Y, Froguel P, Kimura S, Nagai R, Kadowaki T (2003b) Globular adiponectin protected ob/ob mice from diabetes and ApoE-deficient mice from atherosclerosis. J Biol Chem 278:2461-2468.
    Yamauchi T, Kamon J, Waki H, Terauchi Y, Kubota N, Hara K, Mori Y, Ide T, Murakami K, Tsuboyama-Kasaoka N, Ezaki O, Akanuma Y, Gavrilova O, Vinson C, Reitman ML, Kagechika H, Shudo K, Yoda M, Nakano Y, Tobe K, Nagai R, Kimura S, Tomita M, Froguel P, Kadowaki T (2001) The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity. Nat Med 7:941-946.
    Ziemke F, Mantzoros CS (2010) Adiponectin in insulin resistance: lessons from translational research. Am J Clin Nutr 91:258S-261S.

    下載圖示
    QR CODE