生物分子
电子转移
氧化还原
双层
膜
烟酰胺腺嘌呤二核苷酸
化学
电化学
脂质双层
拉曼光谱
X射线光电子能谱
生物膜
材料科学
纳米技术
光化学
电极
化学工程
无机化学
物理化学
有机化学
NAD+激酶
生物化学
物理
工程类
光学
酶
作者
Wei Ma,Yi‐Lun Ying,Lixia Qin,Zhen Gu,Hao Zhou,Dawei Li,Todd C. Sutherland,Hong‐Yuan Chen,Yi‐Tao Long
出处
期刊:Nature Protocols
[Springer Nature]
日期:2013-02-07
卷期号:8 (3): 439-450
被引量:56
标识
DOI:10.1038/nprot.2013.007
摘要
Here we report a protocol to investigate the electron-transfer processes of redox-active biomolecules in biological membranes by electrochemistry using biomimetic hybrid bilayer membranes (HBMs) assembled on gold electrodes. Redox-active head groups, such as the ubiquinone moiety, are embedded in HBMs that contain target molecules, e.g., nicotinamide adenine dinucleotide (NADH). By using this approach, the electron-transfer processes between redox molecules and target biomolecules are mediated by mimicking the redox cycling processes in a natural membrane. Also included is a procedure for in situ surface-enhanced Raman scattering (SERS) to confirm the electrochemically induced conformational changes of the target biomolecules in the HBMs. In addition, each step in constructing the HBMs is characterized by electrochemical impedance spectroscopy (EIS), high-resolution X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The time required for the entire protocol is ∼12 h, whereas the electrochemical measurement of electron-transfer processes takes less than 1 h to complete.
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