title: Analysis of interactions between SNARE proteins using imaging ellipsometer coupled with microfluidic array.
abstract: 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.
authors.affiliation: 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].
authors.forename: Cai
authors.initials: C
authors.lastname: Qi
authors.affiliation: 1] Institute of Biophysics, Chinese Academy of Sciences, #15, Datun Rd., Beijing, 100101, China [2].
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Institute of Mechanics, Chinese Academy of Sciences, #15, Beisihuan West Rd., Beijing, 100190, China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Renke
authors.initials: R
authors.lastname: Yang
authors.affiliation: Xiang-Ya Medical School, Central South University, #932, Lushan South Rd., Changsha, Hunan, 410083, China.
authors.forename: Yinke
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Institute of Mechanics, Chinese Academy of Sciences, #15, Beisihuan West Rd., Beijing, 100190, China.
authors.forename: Tengfei
authors.initials: T
authors.lastname: Kang
authors.affiliation: Institute of Mechanics, Chinese Academy of Sciences, #15, Beisihuan West Rd., Beijing, 100190, China.
authors.forename: Wenxin
authors.initials: W
authors.lastname: Hao
authors.affiliation: Institute of Mechanics, Chinese Academy of Sciences, #15, Beisihuan West Rd., Beijing, 100190, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Jin
authors.affiliation: Institute of Biophysics, Chinese Academy of Sciences, #15, Datun Rd., Beijing, 100101, China.
authors.forename: Taijiao
authors.initials: T
authors.lastname: Jiang
mesh_terms: D000465:Algorithms; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D050765:Qa-SNARE Proteins / Q000235:genetics / Q000378:metabolism*; D050683:R-SNARE Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012031:Refractometry / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
chemical_list: D050765:Qa-SNARE Proteins; D050683:R-SNARE Proteins; D011994:Recombinant Proteins; C084007:SSO2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C106909:Sec22 protein, S cerevisiae; C107127:YKT6 protein, S cerevisiae