Delforge D, Art M, Gillon B, Dieu M, Delaive E, Raes M, Remacle J
Laboratory of Cellular Biochemistry, Facultés Universitaires N-D de la Paix (FUNDP), Namur, Belgium.
Anal Biochem. 1996 Nov 15;242(2):180-6. doi: 10.1006/abio.1996.0451.
Recent developments in allyl chemistry and palladium solubilization allow automated continuous-flow solid-phase synthesis of cyclic or branched peptides, with specific side-chain cleavage and on-line cyclization. In this paper, we adapted the method to the synthesis of cyclic peptides bearing an anchoring tail on a side chain of the cycle. Side products were obtained with the standard procedure and an additional washing step had to be introduced in the synthesis protocol to remove side products resulting from the palladium allyl cleavage step. The method is illustrated by the automated synthesis of cyclo[-DVal-Arg-Gly-Asp-Glu (-epsilon Ahx-Cys-NH2)-] which contains the Arg-Gly-Asp adhesion motif (RGD) recognized by cellular integrins. The tail of the peptide was designed with a thiol at the carboxylic end to allow subsequent grafting by covalent attachment. Such tailed cyclic peptides can be grafted on different supports for new applications in biomaterial design, cell adhesion assays, affinity chromatography, immunization, vaccine development, ELISA kits, and the building of libraries of conformationally constrained peptides.
烯丙基化学和钯增溶技术的最新进展使得环状或支链肽的自动化连续流固相合成成为可能，能够实现特定侧链切割和在线环化。在本文中，我们将该方法应用于合成在环的侧链上带有锚定尾的环状肽。采用标准程序会产生副产物，因此必须在合成方案中引入额外的洗涤步骤，以去除钯烯丙基切割步骤产生的副产物。通过自动合成含有细胞整合素识别的精氨酸 - 甘氨酸 - 天冬氨酸粘附基序（RGD）的环[-DVal-Arg-Gly-Asp-Glu (-εAhx-Cys-NH2)-]来说明该方法。肽的尾部在羧基末端设计有一个硫醇，以便随后通过共价连接进行接枝。这种带尾的环状肽可以接枝到不同的载体上，用于生物材料设计、细胞粘附测定、亲和色谱、免疫、疫苗开发、酶联免疫吸附测定试剂盒以及构建构象受限肽库等新应用。
