Characterization of the Interactions of the Bacterial Cell Division Regulator MinE
III Acknowledgements IV List of Abbreviations VII List of Figures VIII List of Tables X Chapter 1: Introduction 1 1.1: Bacterial Cell Division 1 1.2: Min Protein Interactions 2 1.3: MinD Structure and Interactions with MinE 5 1.4: MinE Residues Required for MinD Binding 9 1.5: Topological Specificity Function in MinE 11 1.6: A Surprising Structure For Full-length MinE 13 1.7: MinE Interaction with Lipids 16 1.8: Thesis Objectives 17 1.9: Neisseria gonorrhoeae versus Escherichia coli Min Proteins 19 1.10: Kinetics of MinE-Stimulated MinD Activity 21 1.11: CD Spectroscopy to Characterize Structural Changes in MinE 22 1.12: Basic Principles of NMR Spectroscopy 24 1.13: Screening of Lipid-Mimetic Environments by the 15 N HSQC Spectrum 28 Chapter 2: Materials and Methods 32 2.1: Plasmids and Peptides 32 2.2: Amplification and Isolation of Plasmids 32 2.3: Transformation 33 2.4: Bacterial Growth and Overexpression 33 2.5: Purification 34 2.6: SDS-PAGE 35 2.7: Protein Concentration Determination 36 2.8: ATPase Assay 37