The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle:
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| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
München
Verl. Dr. Hut
2009
|
| Ausgabe: | 1. Aufl. |
| Schriftenreihe: | Biochemie
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 203 S. graph. Darst. 210 mm x 148 mm, 327 gr. |
| ISBN: | 9783868532913 |
Internformat
MARC
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| 100 | 1 | |a Schmid, Andreas B. |e Verfasser |0 (DE-588)140293566 |4 aut | |
| 245 | 1 | 0 | |a The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle |c Andreas B. Schmid |
| 250 | |a 1. Aufl. | ||
| 264 | 1 | |a München |b Verl. Dr. Hut |c 2009 | |
| 300 | |a 203 S. |b graph. Darst. |c 210 mm x 148 mm, 327 gr. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 0 | |a Biochemie | |
| 502 | |a Zugl.: München, Techn. Univ., Diss., 2009 | ||
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Datensatz im Suchindex
| _version_ | 1804140986931085312 |
|---|---|
| adam_text | 1. INTRODUCTION 1 1.1 PROTEIN FOLDING 1 1.2 MOLECULAR CHAPERONES 3 1.3
THE MOLECULAR CHAPERONE HSP70 6 1.3.1 STRUCTURAL AND FUNCTIONAL
PROPERTIES OF HSP70 8 1.3.2 ATPASE ACTIVITY OF HSP70 11 1.3.3 MODULATION
OF ATPASE ACTIVITY 13 1.3.4 CHAPERONE ACTIVITY OF HSP70 16 1.4 THE
MOLECULAR CHAPERONE HSP90 18 1.4.1 STRUCTURAL AND FUNCTIONAL PROPERTIES
OF HSP90 19 1.4.2 HSP90 COCHAPERONE COMPLEXES 21 1.4.3 ATPASE ACTIVITY
OF HSP90 22 1.4.4 MODULATION OF ATPASE ACTIVITY 24 1.4.5 CHAPERONE
ACTIVITY OF HSP90 26 1.5 THE HSP70-HSP90 CHAPERONE CYCLE 29 2. OBJECTIVE
34 3. MATERIALS & METHODS 35 3.1 MATERIALS 35 3.1.1 CHEMICALS 35 3.1.2
SIZE MARKERS AND KITS 36 3.1.3 PROTEINS AND ANTIBODIES 36 3.1.4
CHROMATOGRAPHIE MATERIAL 37 3.1.5 MISCELLANEOUS MATERIAL 37 3.1.6
EQUIPMENT 38 3.1.7 COMPUTER SOFTWARE 40 3.2 ORGANISMS AND CULTIVATION 41
3.2.1 STRAINS 41 3.2.2 MEDIA 43 3.2.3 GROWTH AND STORAGE OF E. COLI. 45
3.2.4 GROWTH AND STORAGE OF YEAST CELLS 45 3.2.5 GROWTH AND STORAGE OF
HELA CELLS 45 3.3 WORKING TECHNIQUES IN MOLECULAR BIOLOGY 46
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/999629204 DIGITALISIERT
DURCH 3.3.1 PLASMIDS 46 3.3.2 OLIGONUCLEOTIDES 52 3.3.3 MOLECULAR
BIOLOGICAL SOLUTIONS 57 3.3.4 PREPARATION OF PLASMID DNA FROM COLI 58
3.3.5 SEPARATION OF DNA BY AGAROSE GEL ELECTROPHORESIS 59 3.3.6 DNA
ISOLATION FROM AGAROSE GELS 59 3.3.7 PURIFICATION OF PCR PRODUCTS AND
VECTORS 59 3.3.8 DNA SEQUENCING ANALYSIS 59 3.3.9 TRANSFORMATION OF
E.COLI. 59 3.3.10 TRANSFORMATION OF S. CEREVISIAE 60 3.3.11
TRANSFORMATION OF P. PASTORIS 60 3.3.12 TRANSFECTION OF HELA CELLS 61
3.3.13 PCR AMPLIFICATION 61 3.3.14 DNA DIGEST BY RESTRICTION ENZYMES 62
3.3.15 DEPHOSPHORYLATION OF DNA ENDS 62 3.3.16 LIGATION OF DNA FRAGMENTS
63 3.4 PREPARATIVE METHODS 63 3.4.1 EXPRESSION KINETICS 63 3.4.2 GROWTH,
INDUCTION AND HARVEST OF . COLI. 64 3.4.3 GROWTH, INDUCTION AND HARVEST
OF P. PASTORIS 64 3.4.4 CELL DISRUPTION 64 3.5 METHODS FOR PROTEIN
PURIFICATION 65 3.5.1 AFFINITY CHROMATOGRAPHY 65 3.5.2 ION EXCHANGE
CHROMATOGRAPHY 65 3.5.3 HYDROXYAPATITE CHROMATOGRAPHY 66 3.5.4
HYDROPHOBIE INTERACTION CHROMATOGRAPHY 66 3.5.5 GEL FILTRATION
CHROMATOGRAPHY 66 3.5.6 PROTEIN PRECIPITATION 67 3.5.7 CONCENTRATION OF
PROTEINS 67 3.5.8 PROTEIN DIALYSIS 67 3.5.9 STANDARD PURIFICATION OF
HISE-TAGGED PROTEINS 68 3.5.1 3.5.12 PURIFICATION OF UNTAGGED SSA1 FROM
P. PASTORIS 71 3.6 PROTEIN ANALYTICAL WORK TECHNIQUES 72 3.6.1 SOLUTIONS
FOR PROTEINCHEMICAL METHODS 72 3.6.2 SDS-POLYACRYLAMIDEELECTROPHORESIS
73 3.6.3 DETERMINATION OF THE PROTEIN CONCENTRATION WITH BRADFORD 73
3.6.4 COOMASSIE-STAINING OF SDS-GELS 73 3.6.5 IMMUNOBLOTTING (WESTERN
BLOT) 74 3.6.6 ANALYTICAL GEL FILTRATION 74 3.6.7 ANALYTICAL
ULTRA-CENTRIFUGATION 75 3.6.8 CRYSTALLISATION OF PROTEINS 75 3.7
SPECTROSCOPICAL METHODS 76 3.7.1 UVABSORBTION SPECTROSCOPY 76 3.7.2
FLUORESCENCE SPECTROSCOPY 77 3.7.3 CD SPECTROSCOPY 78 3.8 ACTIVITY
ASSAYS FOR PROTEINS IN VIVO 80 3.8.1 CLONING OF THE STI1/HOP FRAGMENTS
IN EXPRESSION VECTORS 80 3.8.2 STEROID HORMONE RECEPTOR ACTIVITY ASSAY
IN S. CEREVISIAE 82 3.8.3 LUCIFERASE ACTIVITY ASSAY IN S. CEREVISIAE 82
3.8.4 V-SRC ACTIVITY ASSAY IN S. CEREVISIAE 83 3.8.5 GR ACTIVITY ASSAY
IN HELA CELLS 83 3.9 ACTIVITY ASSAYS FOR PROTEINS IN VITRO 84 3.9.1
ATPASE ASSAY WITH RADIOACTIVE Y- 32 P-ATP 84 3.9.2 ATPASE ASSAY WITH AN
ATP-REGENERATING SYSTEM 85 3.9.3 AGGREGATION ASSAY WITH LUCIFERASE 86
3.9.4 REFOLDING ASSAY WITH CHEMICALLY DENATURED LUCIFERASE 86 3.9.5
REFOLDING ASSAY WITH THERMALLY DENATURED LUCIFERASE 87 4 4.1.5 ATPASE
ACTIVITY OF SSA1 94 4.1.6 INFLUENCE OF COCHAPERONES ON THE ATPASE
ACTIVITY OF SSA1 96 4.1.7 REFOLDING ASSAY WITH CHEMICALLY DENATURED
LUCIFERASE 109 4.1.8 DISCUSSION OF THE QUALITY OF SSA1 FROM THE TWO
SOURCES 119 4.2 REFOLDING ASSAY WITH THERMALLY DENATURED LUCIFERASE 120
4.3 ANALYSIS OF STIL FRAGMENTS 129 4.3.1 SELECTED STIL FRAGMENTS FOR
PURIFICATION 129 4.3.2 SECONDARY STRUCTURE OF THE PURIFIED FRAGMENTS 130
4.3.3 THERMAL STABILITY OF THE STIL FRAGMENTS 131 4.3.4 BINDING OF
FRAGMENTS TO SSA1 ORYHSP90 137 4.3.5 MINIMAL STIL FRAGMENT THAT INHIBITS
THE YHSP90 ATPASE 137 4.3.6 MINIMAL STIL FRAGMENT THAT ACCELERATES
LUCIFERASE REFOLDING 139 4.3.7 DISCUSSION OF THE STRUCTURAL ORGANISATION
OF STIL 145 4.4 SHR ACTIVITY IN 5. CEREVISIAE 146 4.4.1 GR ACTIVITY IN
HSP90 COCHAPERONE KNOCK-OUTS 146 4.4.2 SHR ACTIVITY IN WILD-TYPE AND
STUA CELLS 146 4.4.3 GR ACTIVITY DEPENDING ON STIL FRAGMENTS IN STILA
CELLS 147 4.4.4 GR ACTIVITY DEPENDING ON HOP FRAGMENTS IN STIJA CELLS
152 4.4.5 GR ACTIVITY DEPENDING ON STIL FRAGMENTS IN WILD-TYPE CELLS 153
4.4.6 GR ACTIVITY DEPENDING ON HOP FRAGMENTS IN WILD-TYPE CELLS 154
4.4.7 DISCUSSION OF SHR ACTIVITY IN YEAST CELLS 155 4.5
HOP-FRAGMENT-DEPENDENT ACTIVITY OF GR IN HELA CELLS 156 4.5. 4.7.3
DISCUSSION OF V-SRC ACTIVITY IN YEAST CELLS 165 4.8 EXPRESSION OF
STEROID HORMONE RECEPTORS IN COLI AND P. PASTORIS. 165 4.8.1
PURIFICATION OF NATIVE GR-LBD FROM COLI 165 4.8.2 PURIFICATION OF
GR-LBD FROM CO//INCLUSION BODIES 167 4.8.3 PURIFICATION OF GR-LBD FROM
P. PASTORIS 169 4.8.4 DISCUSSION OF THE PURIFICATION STRATEGIES 170 5.
SUMMARY 171 6. ABBREVIATIONS 174 7. LITERATURE 177 8. PUBLICATIONS 201
|
| any_adam_object | 1 |
| author | Schmid, Andreas B. |
| author_GND | (DE-588)140293566 |
| author_facet | Schmid, Andreas B. |
| author_role | aut |
| author_sort | Schmid, Andreas B. |
| author_variant | a b s ab abs |
| building | Verbundindex |
| bvnumber | BV035966492 |
| classification_tum | CHE 806d CHE 820d |
| ctrlnum | (OCoLC)643842770 (DE-599)DNB999629204 |
| discipline | Biologie Chemie |
| edition | 1. Aufl. |
| format | Thesis Book |
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| id | DE-604.BV035966492 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T22:08:42Z |
| institution | BVB |
| isbn | 9783868532913 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-018860584 |
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| physical | 203 S. graph. Darst. 210 mm x 148 mm, 327 gr. |
| publishDate | 2009 |
| publishDateSearch | 2009 |
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| series2 | Biochemie |
| spelling | Schmid, Andreas B. Verfasser (DE-588)140293566 aut The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle Andreas B. Schmid 1. Aufl. München Verl. Dr. Hut 2009 203 S. graph. Darst. 210 mm x 148 mm, 327 gr. txt rdacontent n rdamedia nc rdacarrier Biochemie Zugl.: München, Techn. Univ., Diss., 2009 Bindeproteine (DE-588)4276933-4 gnd rswk-swf Polypeptidketten bindende Proteine (DE-588)4311255-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Polypeptidketten bindende Proteine (DE-588)4311255-9 s Bindeproteine (DE-588)4276933-4 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=018860584&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schmid, Andreas B. The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle Bindeproteine (DE-588)4276933-4 gnd Polypeptidketten bindende Proteine (DE-588)4311255-9 gnd |
| subject_GND | (DE-588)4276933-4 (DE-588)4311255-9 (DE-588)4113937-9 |
| title | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle |
| title_auth | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle |
| title_exact_search | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle |
| title_full | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle Andreas B. Schmid |
| title_fullStr | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle Andreas B. Schmid |
| title_full_unstemmed | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle Andreas B. Schmid |
| title_short | The adaptor protein Sti1/Hop connects the Hsp70 and Hsp90 chaperone cycle |
| title_sort | the adaptor protein sti1 hop connects the hsp70 and hsp90 chaperone cycle |
| topic | Bindeproteine (DE-588)4276933-4 gnd Polypeptidketten bindende Proteine (DE-588)4311255-9 gnd |
| topic_facet | Bindeproteine Polypeptidketten bindende Proteine Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=018860584&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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