Derossi D, Calvet S, Trembleau A, Brunissen A, Chassaing G, Prochiantz A
CNRS URA 1414, Ecole Normale Supérieure, 46 rue d'Ulm, 75230 Paris Cedex 05, France.
J Biol Chem. 1996 Jul 26;271(30):18188-93. doi: 10.1074/jbc.271.30.18188.
We have recently reported that a 16-amino acid long polypeptide corresponding to the third helix of the DNA binding domain (homeodomain) of Antennapedia, a Drosophila transcription factor, is internalized by cells in culture (Derossi, D., Joliot, A. H., Chassaing, G., and Prochiantz, A.(1994) J. Biol. Chem. 269, 10444-10450). The capture of the homeodomain and of its third helix at temperatures below 10 degrees C raised the problem of the mechanism of internalization. The present demonstration, that a reverse helix and a helix composed of D-enantiomers still translocate across biological membranes at 4 and 37 degrees C strongly suggests that the third helix of the homeodomain is internalized by a receptor-independent mechanism. The finding that introducing 1 or 3 prolines in the structure does not hamper internalization also demonstrates that the alpha-helical structure is not necessary. The data presented are compatible with a translocation process based on the establishment of direct interactions with the membrane phospholipids. The third helix of the homeodomain has been used successfully to address biologically active substances to the cytoplasm and nucleus of cells in culture (Théodore, L., Derossi, D., Chassaing, G., Llirbat, B., Kubes, M., Jordan, P., Chneiweiss, H., Godement, P., and Prochiantz, A.(1995) J. Neurosci. 15, 7158-7167). Therefore, in addition to their physiological implications (Prochiantz, A., and Théodore, L.(1995) BioEssays 17, 39-45), the present results open the way to the molecular design of cellular vectors.
我们最近报道，一种与果蝇转录因子触角足蛋白（Antennapedia）的DNA结合结构域（同源结构域）的第三螺旋相对应的16个氨基酸长的多肽，可被培养细胞内化（德罗西，D.，乔利奥，A. H.，沙桑，G.，和普罗西安茨，A.（1994年）《生物化学杂志》269，10444 - 10450）。在低于10摄氏度的温度下同源结构域及其第三螺旋的摄取引发了内化机制的问题。目前的证明表明，一个反向螺旋和一个由D - 对映体组成的螺旋在4摄氏度和37摄氏度时仍能跨生物膜转运，这强烈表明同源结构域的第三螺旋是通过一种不依赖受体的机制内化的。在结构中引入1个或3个脯氨酸并不妨碍内化这一发现也表明α - 螺旋结构并非必需。所呈现的数据与基于与膜磷脂建立直接相互作用的转运过程相一致。同源结构域的第三螺旋已成功用于将生物活性物质递送至培养细胞的细胞质和细胞核（西奥多，L.，德罗西，D.，沙桑，G.，利尔巴特，B.，库贝斯，M.，乔丹，P.，施内韦斯，H.，戈德门特，P.，和普罗西安茨，A.（1995年）《神经科学杂志》15，7158 - 7167）。因此，除了它们的生理学意义（普罗西安茨，A.，和西奥多，L.（1995年）《生物论评》17，39 - 45）之外，目前的结果为细胞载体的分子设计开辟了道路。
