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I Index
I Index of Contents
I INDEX OF CONTENTS .......................................................................................................................... I
I.I INDEX OF FIGURES ............................................................................................................................. II
I.II INDEX OF TABLES ............................................................................................................................. IV
I.III LIST OF ABBREVIATIONS ................................................................................................................... V
I.III.I Monosaccharides – abbreviations and symbols .................................................................. VIII
I.III.II Amino acids – three-letter and one-letter abbreviations ....................................................... IX
II ABSTRACT .......................................................................................................................................... X
II ZUSAMMENFASSUNG .......................................................................................................................... XI
1 INTRODUCTION .................................................................................................................................. 1
1.1 PROTEIN GLYCOSYLATION ................................................................................................................ 1
1.1.1 N-Glycosylation ...................................................................................................................... 3
1.1.2 O-Glycosylation...................................................................................................................... 4
1.1.3 Glycosaminoglycan modification ........................................................................................... 5
1.2 O-MANNOSYLATION ......................................................................................................................... 7
1.2.1 Biosynthesis and structure ..................................................................................................... 7
1.2.2 Elongation of the core mannose ............................................................................................ 9
1.2.3 O-Mannosylated proteins ....................................................................................................... 9
1.2.4 Initiation of O-mannosylation ............................................................................................... 10
1.3 PATHOLOGY OF O-MANNOSYLATION DEFECTS ................................................................................. 10
1.3.1 α-Dystroglycan ..................................................................................................................... 10
1.3.2 Dystroglycanopathy ............................................................................................................. 12
1.4 BRAIN EXTRACELLULAR MATRIX ...................................................................................................... 14
1.4.1 Lecticans .............................................................................................................................. 15
1.4.2 Neurofascin .......................................................................................................................... 17
1.4 AIM OF THIS PROJECT .................................................................................................................... 18
2 MATERIALS AND METHODS ........................................................................................................... 19
2.1 MATERIALS .................................................................................................................................... 19
2.1.1 Buffers and Solutions ........................................................................................................... 19
2.1.2 Protein Standards for Electrophoresis ................................................................................. 21
2.1.3 Antibodies ............................................................................................................................ 21
2.2 METHODS ...................................................................................................................................... 22
2.2.1 Purification of O-mannosylated proteins .............................................................................. 22
2.2.2 Protein identification ............................................................................................................ 24
2.2.3 Analysis of O-glycans and glycopeptides ............................................................................ 25
2.2.4 Recombinant expression and purification of neurocan ....................................................... 26
3 RESULTS ........................................................................................................................................... 27
I