Carrigan Christina N, Imperiali Barbara
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Anal Biochem. 2005 Jun 15;341(2):290-8. doi: 10.1016/j.ab.2005.03.026.
Reversible lipid attachment was investigated as a means to deliver small peptides into cells. Two labile straight chain alkyl motifs were developed: a cysteine dodecane disulfide (Cdd) building block and a tyrosine- or serine-myristate ester. Both moieties are cleaved on cell internalization and are compatible with Fmoc solid phase peptide synthesis. A series of fluorophore-labeled peptides that varied in lipophilic content, net charge, and charge distribution were synthesized. The peptides were screened for cellular uptake efficiency as monitored by fluorescence microscopy. Effective peptide transport is based on a distributed net positive charge introduced as lysine residues at the C and/or N terminus of the peptide and the presence of a hydrophobic domain exhibiting an estimated log P4.0. The incorporation of labile lipid motifs into peptides enhances lipophilic character of the peptides and contributes to cellular uptake with minimal alteration to the native sequence.
研究了可逆脂质附着作为将小肽递送至细胞的一种手段。开发了两种不稳定的直链烷基基序：一种半胱氨酸十二烷二硫键（Cdd）构建块以及一种酪氨酸或丝氨酸肉豆蔻酸酯。这两种基团在细胞内化时均会裂解，并且与芴甲氧羰基（Fmoc）固相肽合成兼容。合成了一系列在亲脂性含量、净电荷和电荷分布方面存在差异的荧光团标记肽。通过荧光显微镜监测筛选这些肽的细胞摄取效率。有效的肽转运基于在肽的C端和/或N端引入赖氨酸残基形成的分布性净正电荷以及存在估计log P为4.0的疏水域。将不稳定脂质基序掺入肽中可增强肽的亲脂性，并在对天然序列进行最小程度改变的情况下促进细胞摄取。
