Coagulation protein FVIII binding to phospholipid membranes investigated by fluorescence correlation spectroscopy [Elektronische Ressource] / vorgelegt von Hanna C. Engelke

Coagulation protein FVIII binding tophospholipid membranes investigated byFluorescence Correlation SpectroscopyHanna C. EngelkeMunchen 2010Coagulation protein FVIII binding tophospholipid membranes investigated byFluorescence Correlation SpectroscopyDissertationan der Fakult at fur Physik derLudwig{Maximilians{Universit at Munc henvorgelegt vonHanna C. Engelkeaus WurzburgErstgutachter: Prof. Dr. Joachim R adlerZweitgutachter: Prof. Dr. Roland NetzTag der mundlic hen Prufung: 11. M arz 2010Contents1 Summary 12 Zusammenfassung 33 Introduction 54 Fluorescence Correlation Spectroscopy 94.1 Principle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94.2 Binding experiments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114.3 Quantitative analysis of absolute values . . . . . . . . . . . . . . . . . . . 124.4 Cross-correlation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144.5 Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Analysis of binding isotherms 175.1 Equilibrium binding constants . . . . . . . . . . . . . . . . . . . . . . . . 175.2 Association and dissociation rates . . . . . . . . . . . . . . . . . . . . . . 225.3 Mechanisms of and in uences on protein-membrane binding . . . . . . . 236 Di usion measurements in crowded, scattering media 256.1 Complex media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 256.2 Scattering e ects .