Sainlos Matthieu, Imperiali Barbara
Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139-4307, USA.
Nat Protoc. 2007;2(12):3201-9. doi: 10.1038/nprot.2007.442.
This protocol presents the peptide incorporation of environment-sensitive fluorophores derived from the dimethylaminophthalimide family. The procedure utilizes anhydride precursors of 4-dimethylaminophthalimide (4-DMAP) or 6-dimethylaminonaphthalimide (6-DMN), whose syntheses are described in a related protocol from these authors. In this protocol, the fluorophores are directly incorporated after solid-phase peptide synthesis (SPPS) via on-resin derivatization of peptides prepared using commercially available diamino acids, which are Alloc-protected on the side-chain amino group. The time required to complete the procedure depends on the size and number of peptides targeted. As an alternative to this approach, the corresponding fluorescent amino acids can be obtained in an Fmoc-protected form for convenient use as building blocks in SPPS. This option is described in a related protocol by these authors.
本方案介绍了源自二甲基氨基邻苯二甲酰亚胺家族的环境敏感荧光团的肽掺入方法。该程序使用4-二甲基氨基邻苯二甲酰亚胺（4-DMAP）或6-二甲基氨基萘二甲酰亚胺（6-DMN）的酸酐前体，其合成方法在这些作者的相关方案中有描述。在本方案中，荧光团在固相肽合成（SPPS）后通过使用市售二氨基酸制备的肽的树脂上衍生化直接掺入，所述二氨基酸的侧链氨基上具有Alloc保护。完成该程序所需的时间取决于目标肽的大小和数量。作为该方法的替代方案，可以获得Fmoc保护形式的相应荧光氨基酸，以便方便地用作SPPS中的构建块。这些作者在相关方案中描述了该选项。
