Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
München
Verl. Dr. Hut
2011
|
| Ausgabe: | 1. Aufl. |
| Schriftenreihe: | Biochemie
|
| Schlagworte: | |
| Online-Zugang: | Inhaltstext Inhaltsverzeichnis |
| Beschreibung: | 221 S. Ill., graph. Darst. 210 mm x 148 mm, 288 g |
| ISBN: | 9783868538434 |
Internformat
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| 245 | 1 | 0 | |a Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans |c Andreas Marc Gaiser |
| 250 | |a 1. Aufl. | ||
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Datensatz im Suchindex
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| adam_text |
IMAGE 1
CONTENT
INTRODUCTION 9
PROTEINS 10
PROTEIN FOLDING 10
CATALYZED PROTEIN FOLDING IN VIVO 13
MOLECULAR CHAPERONES 14
RIBOSOMAL ASSOCIATED PROTEINS 15
THE HSP60 FAMILY 25
THE HSPLOO FAMILY 16
THE SMALL HEAT-SHOCK PROTEINS 18
THE HSP70 FAMILY 18
THE HSP90 FAMILY 22
HSP90 22
MEMBERS OF THE HSP90 FAMILY 22
REGULATION OF HSP90 EXPRESSION 23
DOMAIN STRUCTURE OF HSP90 HOMOLOGS 25
STRUCTURAL COMPOSITION OFRLSPSO 26
THE ATPASE CYCLE OF HSP90 29
HSP90 INHIBITORS 31
MODULATION OF THE HSP90 ATPASE ACTIVITY BY CO-CHAPERONES 33
MODULATION OF HSP90 ACTIVITY BY POST-TRANSCRIPTIONOL MODIFCATIONS 39
HSP90 SUBSTRATES 41
THE NEMATODE CAENORHABDIVS ELEGANS 45
BIOLOGY AND LIFE CYCLE OF C. ELEGANS 45
C. ELEGANS AS A MODEL ORGANISM 49
STRESS RESPONSE AND AGEING IN C. ELEGANS 50
OBJECTIVE 53
MATERIAL AND METHODS 55
MATERIALS 56
CHEMICALS 56
SIZE MARKERS AND KITS 57
PROTEINS AND ANTIBODIES 58
CHROMATOGRAPHIE MATERIAL 58
MISCELLANEOUS MATERIAL 59
EQUIPMENT 59
COMPUTER SOFTWARE. 62
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/1010823469
DIGITALISIERT DURCH
IMAGE 2
ORGANISMS AND CULTIVATION 64
BACTERIAL STRAINS 64
C. ELEGANS STRAINS 65
MEDIA AND SOLUTIONS 65
GROWTH AND STORAGE OF E. COLL 67
WORKINSTECHNIQUES IN MOLECULAR BIOLOGY 67
PLASMIDS AND CONSTRUCTS 67
MOLECULAR BIOLOGICAL SOLUTIONS 69
PREPARATION OF PLASMID DNA FROM E. COLI 70
SEPARATION OF DNA BY AGOROSE GET ELECTROPHORESIS 70
DNA ISOLATION FROM AGAROSE GELS 70
PURIFICATION OF PCR PRODUCTS AND PLOSMIDS 70
DNA SEQUENCING ANALYSIS 70
TRANSFORMATION OF E. COLL 71
PCR AMPLIFICATION 71
DNA DIGEST BY RESTRICTION ENDONUCLEASES 72
DEPHOSPHORYLATION OF DNA ENDS , 73
UGATION OF DNA FRAGMENTS 73
PREPARATIVE METHODS 73
EXPRESSION KINETICS 73
GROWTH AND HARVEST OF E. COLI 74
CELL DISRUPTION 74
METHODS FOR PROTEIN PURIFICATION 74
AFFINITY CHROMATOGRAPHY 75
ION EXCHANGE CHROMATOGRAPHY 75
GEL FILTRATION CHROMATOGRAPHY 76
CONCENTRATION OF PROTEINS 76
PROTEIN DIALYSIS 76
STANDARD PURIFICATION OF HISS-TAGGED PROTEINS 77
WORKING TECHNIQUES IN PROTEIN ANALYTICS 79
SOLUTIONS IN PROTEIN CHEMISTRY 79
SDS-POIYACRYTAMIDE ELECTROPHORESIS 80
PROTEIN CONCENTRATION DETERMINATION WITH BRADFORD 80
COOMASSIE STAINING OF SOS GELS 80
PULL DOWNS FROM WORM EXTRACTS 81
IMMUNOBLOTTING WESTERN BLOT) 82
CROSSLINKING EXPERIMENTS WITH GLUTARALDEHYDE 82
ANALYTICAL GEL FILTRATION CHROMATOGRAPHY 83
IMAGE 3
PROTEIN LABELING 83
ANALYTICAL ULTROCENTRIFUGATION 84
SPECTROSCOPICAL METHODS 86
UV-ABSORPTION SPECTROSCOPY AND DETERMINATION OF PROTEIN CONCENTRATION 86
CD SPECTROSCOPY 87
SURFACE PLASMON RESONANCE SPECTROSCOPY (SPR) 90
ACTIVITY TESTS FOR PROTEINS 92
ATPASE ASSAY WITH AN ATP-REGENERATING SYSTEM 92
METHODS IN C. ELEGANS 93
NEMATODE GROWTH AND CULTIVATION 93
LIFESPAN ASSAY 94
MOTILITY ASSAY 94
GENERATION OF TRANSGENIC LINES 94
RNA INTERFERENCE EXPERIMENTS 96
MICROSCOPY, CONFOCAL MICROSCOPY AND FLUORESCENCE RECOVERY AFTER
PHOTOBLEACHING FRAP) 96
RESULTS AND DISCUSSION 99
BASIC ENZYMATIC PROPERTIES OF C E I E G / V S H S P 9 0 / D A F - 21
100
HOMOLOGY TO S. CEREVISIAE AND H. SAPIENS HSP90 PROTEINS 200
PURIFICATION OF RECOMBINANT HSP90 202
THE OLIGOMERIC STATUS OF HSP90 101
ANALYSIS OF THE ATPASE ACTIVITY OF HSP90 102
DETERMINATION OF THE MLCHAELIS-MENTEN CONSTANT (K M) 203
HEAT-DEPENDENT ATPASE ACTIVITY OF HSP90 103
NUCLEOTIDE-INDUCED CONFORMATIONAL CHANGES IN HSP90 204
INHIBITION OF THE ATPASE OF CEHSP90 BY HSP90 INHIBITORS 105
DISTRIBUTION OFTITC-GA IN C. ELEGANS NEMATODES AND INFLUENCE ON THE
MOTILLTY 206
DISCUSSION OF THE BASIC ENZYMATIC PROPERTIES OF HSP90/DAF-21 FROM C.
ELEGANS 108
THE INTERACTION OF STI1 WITH HSP90 AND HSP70 110
C. ELEGANS STI1 SHOWS A SPECIFIC DOMAIN LOSS 220
DETECTION OFSTIL BY A-STIL POLYCLONAL ANTIBODY IN WORM LYSATES 113
THE TRUNCATED STI1 PROTEIN IS REQUIRED FOR THE DEVELOPMENT OF FERTILITY
114
STI1 BINDS TO HSP90 AND INHIBITS ITS ATPASE ACTIVITY. 127
STI1 BINDS TO HSC70 IN THE ABSENCE OF TPR1 220
BINDING OF HSP90 TO STI1 COMPETES WITH HSC70 BINDING 122
HSP90 KNOCKDOWN LEADS TO STIL-LIKE DEFECTS DURING GONAD DEVELOPMENT 123
DISCUSSION OF THE INTERACTION OF STI1 WITH HSP90 AND H S P 70 127
HSP90/DAF-21 ENSURES MUSCLE MAINTENANCE IN AGEING C. ELEGANS 130
A MUTATION IN HSP90 LEADS TO A SHORTER LIFE SPAN AND MOTILITY DEFECTS IN
NEMATODES 130
IMAGE 4
BIOCHEMICAL ANALYSIS OF E292K-HSP90 132
THE HSP90-PROMOTER DIRECTS INTO MULTIPLE CELL TYPES, INCLUDING MUSCLE
CELLS 235
KNOCKDOWN OF HSP90 AND ITS CO-FACTOR UNC-45 LEADS TO DISTORTION OF
SARCOMERE STRUCTURE 236
REDUCTION OFHSP90 LEVELS INDUCES THE STRESS RESPONSE IN BODY WALL MUSCLE
CELLS 137
A TRANSLATIONAL HSP90::YFP REPORTER CONSTRUCT LOCALIZES TO THE A- AND
L-BAND OF THE MUSCLE
SARCOMERE 138
INFLUENCE OFHSP90 CO-CHAPERONES ON THE HSP90::YFP ASSOCIATION TO MUSCLE
STRUCTURES 142
HSP90::YFP ASSOCIATION TO MUSCULAR ULTRASTRUCTURES IS HIGHLY DYNAMIC 243
DISCUSSION OF THE ROLE OF HSP90 IN MUSCLE ORGANIZATION AND MAINTENANCE
IN C. CLEGANS 145
ANALYSIS OF CDC37*HSP90 COMPLEXES IN RESPONSE TO NUCLEOTIDES AND BINDING
OF FURTHER COFACTORS 148
OLIGOMERIC STATUS OF C. ELEGANS CDC37. 248
CDC37 INHIBITS THE ATPASE ACTIVITY OF C. ELEGANS HSP90 149
ANALYSIS OF CDC37*HSP90 COMPLEX FORMATION BY ANALYTICAL
ULTRACENTRIFUGATION (AUC) 150
CDC37 BINDING IS ENHANCED BY NUCLEOTIDE-INDUCED CONFORMATIONAL CHANGES
152
THE CDC37-HSP90 COMPLEX CAN BE DISRUPTED BY THE HSP90 COFACTORS STI1 AND
CEP23 155
THE CDC37'HSP90 COMPLEX IS NOT AFFECTED BY THE CO-CHAPERONE PPH-5 259
AHOL FORMS TERNARY COMPLEXES WITH CDC37 AND HSP90 IN THE ABSENCE OF
NUCLEOTIDES 262
FULL REPLACEMENT OFCDC37 REQUIRES THE C-DOMAIN OF AHOL AND
CONFORMATIONAL CHANGES WITHIN
HSP90 265
DISCUSSION OF THE EFFECTS OF COFACTOR BINDING ON THE CDC37»HSP90 COMPLEX
FORMATION 169
SUMMARY 175
ABBREVIATIONS 181
REFERENCES 185
PUBLICATIONS 219
ACKNOWLEDGMENTS 220
DECLARATION 221 |
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| author | Gaiser, Andreas Marc |
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| discipline | Biologie Chemie |
| edition | 1. Aufl. |
| format | Thesis Book |
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| isbn | 9783868538434 |
| language | English |
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| spelling | Gaiser, Andreas Marc Verfasser (DE-588)144035758 aut Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans Andreas Marc Gaiser 1. Aufl. München Verl. Dr. Hut 2011 221 S. Ill., graph. Darst. 210 mm x 148 mm, 288 g txt rdacontent n rdamedia nc rdacarrier Biochemie Zugl.: München, Techn. Univ., Diss., 2010 Hitzeschock-Proteine (DE-588)4255487-1 gnd rswk-swf Caenorhabditis elegans (DE-588)4147127-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Caenorhabditis elegans (DE-588)4147127-1 s Hitzeschock-Proteine (DE-588)4255487-1 s DE-604 X:MVB text/html http://deposit.dnb.de/cgi-bin/dokserv?id=3708587&prov=M&dok_var=1&dok_ext=htm Inhaltstext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=022497052&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gaiser, Andreas Marc Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans Hitzeschock-Proteine (DE-588)4255487-1 gnd Caenorhabditis elegans (DE-588)4147127-1 gnd |
| subject_GND | (DE-588)4255487-1 (DE-588)4147127-1 (DE-588)4113937-9 |
| title | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans |
| title_auth | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans |
| title_exact_search | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans |
| title_full | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans Andreas Marc Gaiser |
| title_fullStr | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans Andreas Marc Gaiser |
| title_full_unstemmed | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans Andreas Marc Gaiser |
| title_short | Studies on the molecular chaperone Hsp90 and its regulation by co-chaperones in Caenorhabditis elegans |
| title_sort | studies on the molecular chaperone hsp90 and its regulation by co chaperones in caenorhabditis elegans |
| topic | Hitzeschock-Proteine (DE-588)4255487-1 gnd Caenorhabditis elegans (DE-588)4147127-1 gnd |
| topic_facet | Hitzeschock-Proteine Caenorhabditis elegans Hochschulschrift |
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