The crystal structure of a bacterial lysozyme at atomic resolution [Elektronische Ressource] / von Astrid Rau

The crystal structure of a bacterial lysozyme at atomic resolution Dissertation zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) vorgelegt dem Rat der Chemisch-Geowissenschaftlichen Fakultät der Friedrich-Schiller-Universität Jena von Diplom-Chemikerin Astrid Rau geboren am 01.06.1974 in Saalfeld Gutachter: 1. Prof. Dr. R. Hilgenfeld 2. Prof. Dr. D. Klemm Tag der öffentlichen Verteidigung: 01.06.2005 TABLE OF CONTENTS 1. INTRODUCTION 1 1.1 Milestones in lysozyme research 1 1.2 Definition and classification of lysozymes 5 1.3 Catalytic mechanisms of lysozymes 7 1.4 Chalaropsis-type lysozymes 9 1.5 Cellosyl – a Ch-type lysozyme from Streptomyces coelicolor 12 1.7 Aim of the project 13 2. MATERIALS AND METHODS 14 2.1 Materials 14 2.1.1 Proteins 14 2.1.2 Carbohydrates 14 2.1.3 Chemicals 15 2.1.4 Crystallisation screens 15 2.1.5 Dialysing tools, assays, crystallisation materials and cryo-tools 15 2.1.6 Laboratory equipment and synchrotron facilities 16 2.2 Methods 17 2.2.1 Determination of protein purity 17 2.2.2 Determination of protein concentration 18 2.2.3 Dialysis 18 2.2.4 Sample concentration 18 2.2.5 Crystallisation 18 2.2.6 Heavy atom and polysaccharide soaks 19 2.2.7 Cryocooling 20 2.2.8 Data acquisition and processing 20 2.2.8.