Qi Cai, Zhang Hong, Liu Li, Yang Renke, Kang Tengfei, Hao Wenxin, Jin Gang, Jiang Taijiao
1] Institute of Biophysics, Chinese Academy of Sciences, #15, Datun Rd., Beijing, 100101, China [2] Institute of Equipment Technology, Chinese Academy of Inspection and Quarantine, #3, Gaobeidian North Rd., Beijing, 100123, China [3].
1] Institute of Biophysics, Chinese Academy of Sciences, #15, Datun Rd., Beijing, 100101, China [2].
Sci Rep. 2014 Jun 18;4:5341. doi: 10.1038/srep05341.
The soluble N-ethylmaleimide-sensitive factor attachment receptor (SNARE) proteins are small and abundant membrane-bound proteins, whose specific interactions mediate membrane fusion during cell fusion or cellular trafficking. In this study, we report the use of a label-free method, called imaging ellipsometer to analyze the interactions among three SNAREs, namely Sec22p, Ykt6p and Sso2p. The SNAREs were immobilized on the silicon wafer and then analyzed in a pairwise mode with microfluidic array, leading us to discover the interactions between Ykt6p and Sso2p, Sec22p and Sso2p. Moreover, by using the real-time function of the imaging ellipsometer, we were able to obtain their association constants (K(A)) of about 10(4) M(-1). We argue that the use of imaging ellipsometer coupled with microfluidic device will deepen our understanding of the molecular mechanisms underlying membrane fusion process.
可溶性N - 乙基马来酰亚胺敏感因子附着受体（SNARE）蛋白是一类小而丰富的膜结合蛋白，其特异性相互作用介导细胞融合或细胞运输过程中的膜融合。在本研究中，我们报告了一种无标记方法的应用，即成像椭偏仪，用于分析三种SNARE蛋白（即Sec22p、Ykt6p和Sso2p）之间的相互作用。将SNARE蛋白固定在硅片上，然后通过微流控阵列以两两配对的方式进行分析，从而发现了Ykt6p与Sso2p、Sec22p与Sso2p之间的相互作用。此外，利用成像椭偏仪的实时功能，我们能够获得它们约为10⁴ M⁻¹ 的缔合常数（K(A)）。我们认为，成像椭偏仪与微流控装置的结合使用将加深我们对膜融合过程潜在分子机制的理解。
