The effects of cholesterol lowering agents on the proteome of primary human hepatocytes [Elektronische Ressource] / von Martin Wörner

The effects of cholesterol lowering agents on the proteome of primary human hepatocytes Dissertation zur Erlangung des Grades des Doktors der Naturwissenschaften der Naturwissenschaftlich-Technischen Fakultät III Chemie, Pharmazie, Bio- und Werkstoffwissenschaften der Universität des Saarlandes von Martin Wörner Saarbrücken 2009 Tag des Kolloquiums: 12.07.2010 Dekan: Prof. Dr. Stefan Diebels Berichterstatter: Prof. Dr. Rita Bernhardt Prof. Dr. Elmar Heinzle Vorsitzender: Prof. Dr. Uli Müller Akademischer Mitarbeiter: Dr. Klaus Hollemeyer Index IIIInnnnddddeeeexxxx ABBREVIATIONS I SSUUMMMMAARRYY IIVV SSUUMMMMAARRYY IIVVSUMMARY (GERMAN) V 1 INTRODUCTION 1 1.1 CHOLESTEROL 1 1.1.1 PHYSIOLOGICAL ROLE 1 1.1.2 TRANSPORT 2 1.1.3 BIOSYNTHESIS AND DEGRADATION 3 1.1.4 REGULATION OF BIOSYNTHESIS, TRANSPORT AND DEGRADATION 6 1.1.5 INHIBITORS OF CHOLESTEROL BIOSYNTHESIS 8 1.2 PRIMARY HUMAN HEPATOCYTES 11 11..33 PPRROOTTEEOOMMIICCSS 1122 11..33 PPRROOTTEEOOMMIICCSS 11221.3.1 SAMPLE PREPARATION 12 1.3.2 SAMPLE SEPARATION 13 1.3.3 PROTEIN IDENTIFICATION 17 1.3.4 QUANTITATION 19 1.4 AIM OF THE WORK 22 2 MATERIALS 23 2.1 CHEMICALS 23 2.2 INSTRUMENTS / MATERIALS 25 2.3 BUFFER / SOLUTIONS / WATER 26 33 MMEETTHHOODDSS 2277 33 MMEETTHHOODDSS 22773.1 CELL CULTURE 29 3.1.1 PRIMARY HUMAN HEPATOCYTES 29 3.1.