Cell internalization of the third helix of the antennapedia homeodomain is receptor-independent
Cell internalization of the third helix of the antennapedia homeodomain is receptor-independent
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DOI：
10.1074/jbc.271.30.18188
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发表时间：
1996-07-26
影响因子：
4.8
通讯作者：
Prochiantz, A
中科院分区：
文献类型：
--
作者：
Derossi, D;Calvet, S;Prochiantz, A
我们最近报道了一种16个氨基酸长的多肽，其对应于果蝇转录因子--果蝇足的DNA结合结构域（同源结构域）的第三螺旋，被培养的细胞内化（Derossi，D.，Joliot，A. H、Chassaing，G.，和Prochiantz，A.（1994）J.Biol.Chem.269，10444-10450）。在低于10摄氏度的温度下捕获同源结构域及其第三螺旋提出了内化机制的问题。目前的证明，一个反向螺旋和一个螺旋组成的D-对映异构体仍然在4和37摄氏度跨生物膜易位强烈表明，第三个螺旋的同源结构域是内在的受体非依赖性机制。在结构中引入1或3个脯氨酸不妨碍内化的发现也证明α-螺旋结构不是必需的。所提出的数据是兼容的易位过程的基础上建立的直接相互作用与膜磷脂。同源结构域的第三螺旋已成功地用于将生物活性物质定位到培养物中细胞的细胞质和细胞核（西奥多，L.，Derossi，D.，Chassaing，G.，Llirbat，B.，Kubes，M.，Jordan，P.，Chneiweiss，H.，Godement，P.，和Prochiantz，A.等人（1995）J. Neurosci. 15，7158-7167）。因此，除了它们的生理影响之外（Prochiantz，A.，和西奥多，L.（1995）BioEssays 17，39-45），本结果为细胞载体的分子设计开辟了道路。
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 (Theodore, 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 Theodore, L. (1995) BioEssays 17, 39-45), the present results open the way to the molecular design of cellular vectors.