title: Investigating electron-transfer processes using a biomimetic hybrid bilayer membrane system.
abstract: 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.
authors: State Key Laboratory of Bioreactor Engineering, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, P. R. China.
authors: Wei
authors: W
authors: Ma
authors: Yi-Lun
authors: YL
authors: Ying
authors: Li-Xia
authors: LX
authors: Qin
authors: Zhen
authors: Z
authors: Gu
authors: Hao
authors: H
authors: Zhou
authors: Da-Wei
authors: DW
authors: Li
authors: Todd C
authors: TC
authors: Sutherland
authors: Hong-Yuan
authors: HY
authors: Chen
authors: Yi-Tao
authors: YT
authors: Long
mesh_terms: D032701:Biomimetics / Q000379:methods*; D058266:Dielectric Spectroscopy / Q000379:methods; D004563:Electrochemistry / Q000379:methods; D004566:Electrodes; D004579:Electron Transport; D006046:Gold / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force / Q000379:methods; D009243:NAD / Q000737:chemistry; D010084:Oxidation-Reduction; D056951:Photoelectron Spectroscopy / Q000379:methods; D013059:Spectrum Analysis, Raman / Q000379:methods
chemical_list: D008051:Lipid Bilayers; D008567:Membranes, Artificial; D009243:NAD; D006046:Gold
references: Langmuir. 2010 Jun 15;26(12):9710-9
references: 20394443
references: Langmuir. 2006 Apr 11;22(8):3497-505
references: 16584220
references: J Am Chem Soc. 2011 Aug 17;133(32):12366-9
references: 21774485
references: Biomaterials. 2010 Oct;31(30):7827-35
references: 20659764
references: J Immunol Methods. 2003 Jul;278(1-2):19-32
references: 12957393
references: Science. 2006 Sep 29;313(5795):1948-51
references: 17008528
references: Anal Chem. 1998 Nov 15;70(22):4670-7
references: 9844566
references: Nature. 2005 Sep 29;437(7059):656-63
references: 16193040
references: Biochim Biophys Acta. 1963 Oct 22;70:554-82
references: 14085940
references: Nat Chem. 2010 Nov;2(11):929-936
references: 20966948
references: Nat Protoc. 2007;2(11):2719-27
references: 18007607
references: Langmuir. 2010 Nov 16;26(22):17674-8
references: 20925377
references: J Am Chem Soc. 2003 Oct 15;125(41):12457-63
references: 14531689
references: Nat Protoc. 2006;1(1):324-36
references: 17406253
references: Langmuir. 2008 Aug 19;24(16):8785-93
references: 18616308
references: J Am Chem Soc. 2007 Dec 26;129(51):16249-56
references: 18052068
references: J Am Chem Soc. 2007 Feb 21;129(7):2094-100
references: 17263532
references: Free Radic Biol Med. 1988;4(1):51-72
references: 2830175
references: Nat Protoc. 2010 Jun;5(6):1096-106
references: 20539285
references: J Am Chem Soc. 2008 Apr 23;130(16):5579-85
references: 18373346
references: Biophys J. 1998 Oct;75(4):1874-85
references: 9746528
references: Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):10209-14
references: 12119384
references: Nat Protoc. 2011 Apr;6(4):523-39
references: 21455188
references: Annu Rev Biochem. 1994;63:675-716
references: 7979252
references: J Am Chem Soc. 2006 Feb 8;128(5):1711-6
references: 16448146
references: Nature. 1998 Apr 16;392(6677):677-84
references: 9565029
references: J Phys Chem B. 2006 Mar 9;110(9):4472-9
references: 16509751
references: Nat Protoc. 2008;3(10):1654-9
references: 18833202
references: Science. 1996 Jan 5;271(5245):43-8
references: 8539599
references: Cell. 2005 Jul 29;122(2):221-33
references: 16051147
references: Antioxid Redox Signal. 2009 Apr;11(4):861-905
references: 19239350
references: J Am Chem Soc. 2004 Oct 20;126(41):13363-76
references: 15479093
references: Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6420-3
references: 8393566
references: Plant Physiol. 2011 Apr;155(4):1477-85
references: 21205617
references: Acc Chem Res. 2002 Mar;35(3):149-57
references: 11900518
references: Langmuir. 2004 Jan 6;20(1):175-80
references: 15745017
references: Science. 2006 Mar 10;311(5766):1430-6
references: 16469879
references: Langmuir. 2007 Jul 3;23(14):7666-71
references: 17539668
references: Angew Chem Int Ed Engl. 2002 Jun 3;41(11):1828-52
references: 19750613