Enzymatic Catalyzed Synthesis and Triggered Gelation of Ionic Peptides
Enzymatic Catalyzed Synthesis and Triggered Gelation of Ionic Peptides
复制标题
离子肽的酶催化合成和触发凝胶化
DOI：
10.1021/la100623y
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发表时间：
2010-07-06
期刊：
影响因子：
3.9
通讯作者：
Miller, Aline F.
中科院分区：
文献类型：
--
作者：
Guilbaud, Jean-Baptiste;Vey, Elisabeth;Miller, Aline F.
我们研究了使用蛋白酶嗜热菌蛋白酶驱动非胶凝前体合成和胶凝离子互补肽的可能性。在该系统中，短肽片段不断相互转化，形成动态肽库，最终有利于通过分子自组装合成热力学稳定的肽。将固定浓度(0.3 mg mL(-1))的嗜热菌蛋白酶添加到短四肽FEFK的溶液(0-300 mg mL(-1))中。最初，蛋白酶将所有样品中的四肽部分水解成二肽。随后，发现通过反向水解形成了更长的肽序列。发现不同序列的稳定性取决于它们的自组装特性。自组装成富含反平行β-折叠纤维的序列成为逆水解反应的稳定产物，而形成的其他序列则不稳定并随着孵育时间的增加而消失。最终，该系统的主要产物是八肽，这表明它代表了该动态库的热力学有利产物。其浓度决定了样品的凝胶化行为，并且根据四肽的初始浓度获得模量高达 25 k Pa 的凝胶。
We investigate the possibility of using the protease thermolysin to drive the synthesis and gelation of ionic-complementary peptides from nongelling precursors. In this system, short peptide fragments are continuously interconverted to form a dynamic peptide library, which eventually favors synthesis of peptides that are thermodynamically stabilized by molecular self-assembly. Thermolysin was added at a fixed concentration (0.3 mg mL(-1)) to solutions (0-300 mg mL(-1)) of the short tetrapeptide FEFK. Initially, the protease partially hydrolyzed the tetrapeptide into dipeptides in all samples. Subsequently, longer peptide sequences were found to form through reverse-hydrolysis. The stability of the different sequences was found to be dependent on their self-assembling properties. The sequences that self-assembled into antiparallel beta-sheet rich Fibers became the stable products for the reverse hydrolysis reaction, while the others formed were unstable and disappeared with increasing incubation time. Ultimately, the main product of the system was octapeptide, which suggests that it represents the thermodynamically favored product of this dynamic library. Its concentration dictated the gelation behavior of the sample, and gels with moduli up to 25 k Pa where obtained depending on the initial concentration of tetrapeptide.