|Budget Amount *help
¥1,600,000 (Direct Cost : ¥1,600,000)
Fiscal Year 1992 : ¥300,000 (Direct Cost : ¥300,000)
Fiscal Year 1991 : ¥1,300,000 (Direct Cost : ¥1,300,000)
Intercellular communication is mediated in many tissues by a major intrinsic protein called gap junction. It has been suggested, from analysis of the sequence of the gap junction protein, that it traverses the lipid bilayer via a transmembrane amphiphilic helix, which is essential for the formation of an intercellular aqueous channel. In the present study, we applied "Monolayer reaction method" to the preparation of the polypeptide composed of two amphiphilic helices jointed with a hydrophilic spacer between them, and examined its activity for the formation of an inter-vesicular aqueous channel as a gap junction model.
The starting material, MPrM, was obtained by polymerization of the N-carboxy anhydride of L-glutamic acid eta-methyl ester with 1,3-diamino 2-propanohol as an initiator in 1,2-dichloroethane. And then MPrM was saponified by using the monolayer reaction method to give the amphiphilic nature to MPrM, am-MPrM. It can be concluded, from CD spectra of am-MPrM in dipalmitoylphosphatidylcholine, gel filtration analysis and inter-vesicular transport of D_2O, that am-MPrM incorporates its two amphiphilic helices to the bilayer membrane of different vesicles, respectively, to induce the vesicular adhesion and then associates each other in the membranes to form an aqueous channel, through which the micro-injection of D_2O can be effectively made. The vesicular recognition ability of the polypeptide will be required to make more realistic model systems.