School of Materials Science and Engineering and Centre for Biomimetic Sensor Science , Nanyang Technological University , 50 Nanyang Drive , Singapore 637553 , Singapore.
Department of Chemistry and Chemical Engineering , Chalmers University of Technology , 41296 Göteborg , Sweden.
Anal Chem. 2018 Jun 19;90(12):7458-7466. doi: 10.1021/acs.analchem.8b00974. Epub 2018 Jun 8.
Nanoplasmonic sensors have emerged as a promising measurement approach to track biomacromolecular interactions involving lipid membrane interfaces. By taking advantage of nanoscale fabrication capabilities, it is possible to design sensing platforms with various architectural configurations. Such capabilities open the door to fabricating lipid membrane-coated nanoplasmonic sensors with varying degrees of membrane curvature in order to understand how biomacromolecular interaction processes are influenced by membrane curvature. Herein, we employed an indirect nanoplasmonic sensing approach to characterize the fabrication of supported lipid bilayers (SLBs) on silica-coated nanowell and nanodisk sensing platforms and to investigate how membrane curvature influences membrane-peptide interactions by evaluating the corresponding measurement responses from different spectral signatures that are sensitive to specific regions of the sensor geometries. SLBs were prepared by the vesicle fusion method, as monitored in real-time by nanoplasmonic sensing measurements and further characterized by fluorescence recovery after photobleaching (FRAP) experiments. By resolving different spectral signatures in the nanoplasmonic sensing measurements, it was determined that peptide binding induces membrane disruption at positively curved membrane regions, while peptide binding without subsequent disruption was observed at planar and negatively curved regions. These findings are consistent with the peptide's known preference to selectively form pores in positively curved membranes, providing validation to the nanoplasmonic sensing approach and highlighting how the integration of nanoplasmonic sensors with different nanoscale architectures can be utilized to study the influence of membrane curvature on biomacromolecular interaction processes.
纳米等离子体传感器已经成为一种很有前途的测量方法,可以跟踪涉及脂质膜界面的生物大分子相互作用。利用纳米级制造能力,可以设计具有各种结构配置的传感平台。这种能力为制造具有不同程度膜曲率的脂质膜涂层纳米等离子体传感器开辟了道路,以便了解生物大分子相互作用过程如何受到膜曲率的影响。在此,我们采用间接纳米等离子体传感方法来表征在涂有二氧化硅的纳米井和纳米盘传感平台上制备的支撑脂质双层(SLB),并通过评估来自不同光谱特征的相应测量响应来研究膜曲率如何影响膜-肽相互作用,这些光谱特征对传感器几何形状的特定区域敏感。通过囊泡融合方法制备 SLB,并通过纳米等离子体传感测量进行实时监测,并用荧光恢复后漂白(FRAP)实验进一步表征。通过解析纳米等离子体传感测量中的不同光谱特征,确定肽结合在正曲率膜区域诱导膜破裂,而在平面和负曲率区域观察到肽结合而没有随后的破裂。这些发现与肽选择性地在正曲率膜中形成孔的已知偏好一致,为纳米等离子体传感方法提供了验证,并强调了如何将具有不同纳米级结构的纳米等离子体传感器与不同纳米级结构的纳米等离子体传感器集成,以研究膜曲率对生物大分子相互作用过程的影响。