Microwave Assisted SPPS of Amylin and Its Toxicity of the Pure Product to RIN-5F Cells
Microwave Assisted SPPS of Amylin and Its Toxicity of the Pure Product to RIN-5F Cells
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DOI：
10.1002/bip.21370
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发表时间：
2010-01-01
期刊：
影响因子：
2.9
通讯作者：
Govender, Thavendran
中科院分区：
文献类型：
--
作者：
Muthusamy, Karen;Albericio, Fernando;Govender, Thavendran
胰岛淀粉样多肽（IAPP）或胰淀素是一种含有37个氨基酸的多肽，在II型糖尿病患者的胰岛中以淀粉样蛋白聚集体的形式存在。本文报道了微波辅助固相多肽合成胰淀素的新方法。最有效的合成使用双重和三重偶联和10当量。的氨基酸。双偶联用于大多数氨基酸，而三重偶联用于选定区域的氨基酸。发现在胰淀素中形成二硫键的最有效方法是碘氧化。当在二硫键形成之前用HPLC纯化粗肽时，获得最高纯度的胰淀素。测定合成的胰淀素产物对RIN-5 F细胞的细胞毒性。合成的胰淀素在浓度>35 μ M时表现出细胞毒性的指数增加。胰淀素样品的透射电子显微镜研究表明，当45 μ M合成胰淀素溶液在合适的缓冲液中孵育6小时时，自发形成不溶性淀粉样蛋白原纤维。(C)2010 Wiley Periodicals，Inc. Biopolymers（Pept Sci）94：323-330，2010.
The 37-amino acid polypeptide islet amyloid polypeptide (IAPP), or amylin, is found as amyloid aggregates in the islets of Langerhans in patients with type II diabetes. Herein, we report an efficient microwave assisted solid phase peptide synthesis of amylin (IAPP). The most efficient synthesis used double and triple couplings and 10 equiv. of amino acids. Double couplings were used for most amino acids, whereas triple couplings were utilized for amino acids in selected regions. The most effective method for formation of the disulfide bond in amylin was found to be iodine oxidation. The highest purity amylin was obtained when the crude peptide was purified with HPLC before formation of the disulfide bond. The cytotoxicity of the synthesized amylin product to RIN-5F cells was determined. The synthesized amylin exhibits an exponential increase of cytotoxicity at concentrations >35 mu M. Transmission electron microscope studies of a sample of amylin shows that insoluble amyloid fibrils spontaneously formed when 45 mu M solution of synthesized amylin was incubated in a suitable buffer for 6 h. (C) 2010 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 94: 323-330, 2010.