Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
Nat Protoc. 2011 Jun;6(6):761-71. doi: 10.1038/nprot.2011.324. Epub 2011 May 12.
This protocol provides a detailed procedure for the preparation of stapled α-helical peptides, which have proven their potential as useful molecular probes and as next-generation therapeutics. Two crucial features of this protocol are (i) the construction of peptide substrates containing hindered α-methyl, α-alkenyl amino acids and (ii) the ring-closing olefin metathesis (RCM) of the resulting resin-bound peptide substrates. The stapling systems described in this protocol, namely bridging one or two turns of an α-helix, are highly adaptable to most peptide sequences, resulting in favorable RCM kinetics, helix stabilization and promotion of cellular uptake.
本方案提供了一种制备订书肽的详细步骤，该肽已被证明具有作为有用的分子探针和下一代治疗剂的潜力。本方案的两个关键特点是：(i) 构建含有受阻 α-甲基、α-烯基氨基酸的肽底物，以及 (ii) 所得树脂结合肽底物的闭环烯烃复分解 (RCM)。本方案中描述的订书系统，即桥连一个或两个α-螺旋圈，高度适应大多数肽序列，从而产生有利的 RCM 动力学、螺旋稳定和促进细胞摄取。
