Spectro-Electrochemical Investigations on Immobilised Heme Proteins [Elektronische Ressource] / Murat Sezer. Betreuer: Peter Hildebrandt

Murat Halstead

Informations

• Publié par: technische_universitat_berlin
• Publié le: 01 janvier 2011
• Nombre de lectures: 16
• Langue: English
• Poids de l'ouvrage: 5 Mo

Extrait

Spectro-Electrochemical Investigations on
Immobilised Heme Proteins


vorgelegt von
Diplom Chemiker
Murat Sezer
aus Berlin


von der Fakultät II – Mathematik und Naturwissenschaften
der Technischen Universität Berlin
zur Erlangung des Akademischen Grades
Doktor der Naturwissenschaften
Dr. rer. nat.

genehmigte Dissertation


Promotionsausschuss:

Vorsitzender: Prof. Dr. Andreas Grohmann
Berichter: Prof. Dr. Peter Hildebrandt
Berichterin: Prof. Dr. Ulla Wollenberger


Tag der wissenschaftlichen Aussprache: 20. Juli 2011


Berlin 2011

D83






































Note

Except for chapter 7.3, this thesis is written in British English. Chapter 7.3 is written
in American English since it is a manuscript that has been accepted for publication
in an American journal (J. Phys. Chem. B.). Moreover, the references in chapter 7.3
are listed separately in the end of chapter 7.3. Chapters 5.4 and 6.3 have been
published in Phys. Chem. Chem. Phys. The references in chapters 5.4 and 6.4 are
also listed seperately in the end of the respective chapters. All other references are
listed in chapter 10.





















Table of Contents

Abbreviations........................................................................................................................I

Symbols...............................................................................................................................II

Physical constants ..............................................................................................................III

Abstract............................................................................................................................... V
Zusammenfassung ............................................................................................................ IX


1 Introduction.....................................................................................................................1

2 Theoretical Background.................................................................................................5
2.1 Raman Spectroscopy ............................................................................................................5
2.1.1 Normal Modes.................................................................................................................................. 5
2.1.2 Raman Scattering ............................................................................................................................ 6
2.1.3 Raman Signal Intensity.................................................................................................................... 7
2.1.4 Polarisability Tensor Components.................................................................................................... 8
2.2 Resonance Raman Effect......................................................................................................9
2.3 Surface Enhanced Raman Spectroscopy ............................................................................ 12
2.4 Electrochemical Interface at the Bare Silver Electrode ........................................................ 14
2.5 Self Assembled Monolayers of Alkanethiol Derivatives........................................................ 15
2.6 Electron Transfer Theory..................................................................................................... 17
2.6.1 Homogeneous Electron Transfer ................................................................................................... 18
2.6.2 Heterogeneous Electron Transfer.................................................................................................. 21
2.7 Heme Proteins .................................................................................................................... 23

3 Instrumentation.............................................................................................................25
3.1 Excitation Source ................................................................................................................ 25
3.2 Sample Arrangement........................................................................................................... 26
3.3 Electrochemical Setup......................................................................................................... 28
3.4 Spectroscopic and Spectro<Electrochemical Setup.............................................................. 28

4 Methods.........................................................................................................................31
4.1 Resonance Raman Spectroscopy of Heme Proteins ........................................................... 31
4.2 Surface Enhanced Resonance Raman Spectroscopy of Heme Proteins ............................. 34
4.2.1 Electrode preparation..................................................................................................................... 34
4.2.3 Time Resolved Potential Controlled Measurements...................................................................... 36
4.2.4 SERR Spectroscopic Study of Catalytic Oxidation Processes ...................................................... 37
4.3 Component Fit Analysis....................................................................................................... 38
4.4 Protein Film Voltammetry .................................................................................................... 40
4.4.1 Protein Film Voltammetry under non<Turnover Conditions............................................................ 41
4.4.2 Protein Film Voltammetry under Turnover Conditions ................................................................... 44
4.5 Computational Chemistry .................................................................................................... 45

5 Silver-Gold Hybrid Devices for Spectro-Electrochemistry........................................47
5.1 Analytical Model for the Optical Performance ...................................................................... 48
5.2 Cytochrome c...................................................................................................................... 51
5.3 Overpotential Dependence of Heterogeneous Electron Transfer Rates............................... 52

5.4 Multi<layer electron transfer across nanostructured Ag<SAM<Au<SAM junctions probed
by surface enhanced Raman spectroscopy..................................................................55


6 Human Sulphite Oxidase..............................................................................................65
6.1 Structure of Human Sulphite Oxidase.................................................................................. 66
6.2 Catalytic Mechanism of Sulphite Oxidases.......................................................................... 66

6.3 Redox properties and catalytic activity of surface<bound human sulfite oxidase studied
by a combined surface enhanced resonance Raman spectroscopic and
electrochemical approach.............................................................................................69

7 Membrane Bound Hydrogenase from Ralstonia eutropha H16 ................................85
7.1 Structure of the Membrane Bound Hydrogenase from Ralstonia eutropha H16................... 86
7.2 Redox Chemistry and Catalytic Cycle.................................................................................. 88

7.3 Role of the HoxZ subunit in the electron transfer pathway of the membrane<bound
[NiFe]<hydrogenase from Ralstonia eutropha H16 immobilized on electrodes.............91

8 Other Projects .............................................................................................................115
8.1 Surface Enhanced Resonance Raman Spectroscopic Investigations on Xenobiotic
Reductase A....................................................................................................................... 115
8.1.1 RR and SERR Spectra Using 8<Aminooctanethiol Coating..........................................................116
8.1.2 SERR Spectra Using 6<Aminohexanethiol Coating......................................................................118
8.2 Resonance Raman Spectroscopic Investigations of Mitochondria Preparations from Mouse
Pancreas............................................................................................................................ 120

9 Conclusion and Outlook ............................................................................................123
9.1 Silver<Gold Hybrid Devices............................................................................................... 123
9.2 Human Sulphite Oxidase....................................