Synthesis of poly(Pro-Hyp-Gly)n by direct polycondensation of (Pro-Hyp-Gly)n, where n = 1, 5, and 10, and stability of the triple-helical structure
Synthesis of poly(Pro-Hyp-Gly)n by direct polycondensation of (Pro-Hyp-Gly)n, where n = 1, 5, and 10, and stability of the triple-helical structure
复制标题
DOI：
10.1002/bip.20348
复制
发表时间：
2005-10-15
期刊：
影响因子：
2.9
通讯作者：
Tanihara, M
中科院分区：
文献类型：
--
作者：
Kishimoto, T;Morihara, Y;Tanihara, M
Pro-Hyp-Gly是纤维性胶原蛋白中发现的一种特有的氨基酸序列，(Pro-Hyp-Gly)(10)是一种稳定的三螺旋结构，被广泛用作胶原模型肽。在这里，我们用1-羟基苯并三氮唑和1-ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide盐酸盐在磷酸盐缓冲液(pH=7.4)和二甲基亚砜(DMSO)溶液中，在20℃下反应48h，通过直接缩聚(Pro-Hyp-Gly)(N)，其中n=1，5和10，合成了由Pro-Hyp-Gly序列组成的多肽。(Pro-Hyp-Gly)(5)和(Pro-Hyp-Gly)(10)在DMSO中反应成功地得到了分子量超过10,000的多肽，在pH=7.4的磷酸盐缓冲液中反应得到低相对分子质量的产物。相反，在pH=7.4的磷酸盐缓冲液中，当Pro-Hyp-Gly的浓度为50 mg/mL时，可得到相对分子质量大于10,000的多肽。傅立叶变换红外光谱(FTIR)和核磁共振氢谱(H-1)表明，本文所述的(Pro-Hyp-Gly)(10)的聚合反应没有副反应。所得多肽均呈现类胶原蛋白的三螺旋结构，聚(Pro-Hyp-Gly)和聚(Pro-Hyp-Gly)(10)的三螺旋结构在T=80℃时保持稳定，这表明除了高Hyp含量外，高相对分子质量还促进了三螺旋结构的稳定性。此外，透射电子显微镜观察表明，聚(ProHyp-Gly)(10)聚集形成宽度约为10 nm的类纳米纤维结构，这表明Pro-Hyp-Gly重复序列包含了足够的信息，可以形成三螺旋结构，以及随后的类纳米纤维结构形成。(C)2005年威利期刊公司。
Pro-Hyp-Gly is a characteristic amino acid sequence found in fibrous collagens, and (Pro-Hyp-Gly)(10), which has been widely used as a collagen-model peptide, forms a stable triple-helical structure. Here, we synthesized polypeptides consisting of the Pro-Hyp-Gly sequence by direct polycondensation of (Pro-Hyp-Gly)(n), where n = 1, 5, and 10, using 1-hydroxybenzotriazole and 1-ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide hydrochloride in both phosphate buffer (pH = 7.4) and dimethylsulfoxide (DMSO) solutions for 48 h at 20 degrees C. The reaction of (Pro-Hyp-Gly)(5) and (Pro-Hyp-Gly)(10) in DMSO successfully gave polypeptides with molecular weights over 10,000, whereas low molecular weight products were obtained by reaction in phosphate buffer (pH = 7.4). In contrast, Pro-Hyp-Gly at a concentration of 50 mg/mL in phosphate buffer (pH = 7.4) gave polypeptides with molecular weights over 10,000. The Fourier transform infrared (FTIR) and H-1 nuclear magnetic resonance (NMR) spectra of poly(Pro-Hyp-Gly)(10) revealed that the polymerization of (Pro-Hyp-Gly)(10) described in this report had no side reactions. Each polypeptide obtained shows a collagen-like triple-helical structure, and the triple-helical structures of poly(Pro-Hyp-Gly) and poly(Pro-Hyp-Gly)(10) were stable up to T = 80 degrees C, which suggests that the high molecular weight promotes stability of the triple-helical structure, in addition to the high Hyp content. Furthermore, transmission electron microscopy (TEM) observations show that poly(ProHyp-Gly)(10) aggregates to form nanofiber-like structures about 10 nm in width, which suggests that a Pro-Hyp-Gly repeating sequence contains enough information for triple-helix formation, and for subsequent nanofiber-like structure formation. (c) 2005 Wiley Periodicals, Inc.