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.
[http://dx.doi.org/10.1038/srep05341]
This study was supported by Project 973 (2014CB910500), National Natural Science Foundation of China (31300632), National Science and Technology Major Project of the Ministry of Science and Technology of China (2013ZX10004611-002), The National Basic Research Program (Project 973) of China (2009CB320300), The National Basic Research Program of China (2009CB320302), The National High Technology Research Development Program of China (2008AA02Z419), The International Science & Technology Cooperation Program of China (2012DFG31880), Projects of Sino-Portugal S&T Cooperation 2010-2015, The National Natural Science Foundation of China (21305147), The Instrument Developing Project of the Chinese Academy of Sciences (KJCX2-YW-M04, KJCX2-YW-M03), Basic Scientific Research Special Foundation of Chinese Academy of Inspection and Quarantine (2010JK002) and China Postdoctoral Science Foundation funded project (20080440533).
The article was published as:
Qi C,Zhang H,Liu L,et al. Analysis Of Interactions Between Snare Proteins Using Imaging Ellipsometer Coupled With Microfluidic Array[J].Scientific Reports,2014,4:5341.