Sakiyama H, Tadokoro S, Nakanishi M, Hirashima N
Nagoya City University, Mizuho-ku, Japan.
Inflamm Res. 2009 Mar;58(3):139-42. doi: 10.1007/s00011-008-7173-z.
We monitored the membrane fusion of liposomes to determine if the minimal components of soluble N-ethyl maleimide-sensitive factor attachment protein receptor (SNARE), which is involved in mast cell exocytosis, have fusogenic activity.
Three core components of SNARE were reconstituted into liposomes. Membrane fusion between liposomes containing vesicle associated membrane protein (VAMP) -7 or -8 and liposomes containing synaptosomal-associated protein 23 kDa (SNAP23) and syntaxin-3 or -4 was monitored by fluorescence resonance energy transfer.
The combination of SNAP23/syntaxin-3/VAMP-8 showed the most efficient liposome-liposome fusogenic activity. Liposomes with VAMP-7 exhibited poor fusogenic activity regardless of the syntaxin isoform.
The core components of SNAP23, syntaxin-3, and VAMP-8 appear to be minimal machinery to induce membrane fusion, while VAMP-7 appears to be unessential for membrane fusion.
我们监测脂质体的膜融合,以确定参与肥大细胞胞吐作用的可溶性N - 乙基马来酰亚胺敏感因子附着蛋白受体(SNARE)的最小组成部分是否具有融合活性。
将SNARE的三个核心组分重组到脂质体中。通过荧光共振能量转移监测含有囊泡相关膜蛋白(VAMP)-7或-8的脂质体与含有23 kDa突触体相关蛋白(SNAP23)和 syntaxin-3或-4的脂质体之间的膜融合。
SNAP23/syntaxin-3/VAMP-8的组合表现出最有效的脂质体 - 脂质体融合活性。无论syntaxin异构体如何,含有VAMP-7的脂质体均表现出较差的融合活性。
SNAP23、syntaxin-3和VAMP-8的核心组分似乎是诱导膜融合的最小机制,而VAMP-7似乎对膜融合并非必需。