Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition: = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. und engl. Sprache |
| Beschreibung: | VI, 156 S. Ill., graph. Darst. |
Internformat
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Datensatz im Suchindex
| _version_ | 1804151984139272192 |
|---|---|
| adam_text | CONTENTS
ABBREVIATION INDEX V
ZUSAMMENFASSUNG 1
ABSTRACT 2
1 INTRODUCTION _ 3
1.1 SERINE PROTEASE INHIBITOR (SERPIN) SUPERFAMILY 4
1.2 CORTICOSTEROID-BINDING GLOBULIN IN STEROID BINDING AND RELEASE 7
1.3 PROTEIN DESIGN APPROACHES 10
1.4 AIMS OF THE PRESENT WORK 10
2 MATERIALS AND METHODS 13
2.1 MATERIALS 13
2.1.1 CHEMICALS 13
2.1.2 SOLUTIONS FOR POLYACRYLAMIDE GEL ELECTROPHORESIS 14
2.1.3
E. COLI
STRAINS 15
2.1.4 ENZYMES 15
2.1.5 PLASMIDS AND PCR-PRIMEIS 15
2.2 METHODS 19
2.2.1 CONSTRUCT DESIGN 19
2.2.2 MUTAGENESIS 20
2.2.3 TRANSFORMATION OF CELLS 20
2.2.4 PROTEIN OVEREXPRESSION AND PURIFICATION OF CBG 20
2.2.5 PROTEIN OVEREXPRESSION AND PURIFICATION OF ACT-BASED DESIGNS ...
21
2.2.6 REACTIVE CENTER LOOP CLEAVAGE ASSAYS 21
2.2.7 CIRCULAR DICLIROISM SPECTROSCOPY 22
2.2.8 POLYMERIZATION ASSAY USING NATIVE PAGE 23
2.2.9 ISOTHERMAL TITRATION CALORIMETRY 23
2.2.10 REDUCTIVE METHYLATION OF LYSINES 24
2.2.11 CRYSTALLIZATION 24
2.2.12 MUMBO CALCULATIONS 26
2.2.13 CRYSTALLOGRAPHIC DATA COLLECTION, SOLUTION OF THE PHASE PROBLEM
AND
MODEL REFINEMENT 28
2.2.14 DATA QUALITY INDICATORS - 29
2.2.15 MODEL QUALITY AND ANALYSIS 29
2.2.16 HOMOLOGY MODELLING 30
2.2.17 SOFTWARE FOR DATA PRESENTATION 30
III
HTTP://D-NB.INFO/1045597201
CONTENTS
3 RESULTS 31
3.1 CBG 31
3.1.1 MUTAGENESIS, EXPRESSION AND PURIFICATION 31
3.1.2 REACTIVE CENTER LOOP CLEAVAGE 37
3.1.3 CIRCULAR DICLIROISM SPECTROSCOPY AND THERMAL STABILITY STUDIES ...
38
3.1.4 ISOTHERMAL TITRATION CALORIMETIY 42
3.1.5 CRYSTALLIZATION AND DATA COLLECTION OF HUMAN CBG 44
3.1.6 STRUCTURE OF CLEAVED HUMAN CBG IN COMPLEX WITH PROGESTERONE . . 46
3.1.7 HOMOLOGY MODELS OF HUMAN CBG AND COMPARISON OF HUMAN AND
RAT CBG 51
3.2 ACT-BASED DESIGNS 54
3.2.1 MUTAGENESIS, EXPRESSION AND PURIFICATION 54
3.2.2 REACTIVE CENTER LOOP CLEAVAGE 60
3.2.3 CIRCULAR DICHROISM SPECTROSCOPY AND THERMAL STABILITY STUDIES . .
. 62^
3.2.4 ISOTHERMAL TITRATION CALORIMETRY 65
3.2.5 CRYSTALLIZATION AND DATA COLLECTION OF METHYLATED NATIVE DBG-1 . .
71
3.2.6 STRUCTURE OF METHYLATED NATIVE DBG-1 74
3.2.7 CIYSTALLIZATION AND DATA COLLECTION OF CLEAVED ACT-BASED DESIGNS .
77
3.2.8 STRUCTURAL COMPARISON OF CLEAVED ACT DESIGNS 83
3.2.9 STRUCTURE OF CDBG-NB IN A STABILIZED CLOSED STATE COMPARED TO
MUMBO CALCULATIONS 86
3.2.10 HYDROCORTISONE BINDING DETERMINANTS IN ACT DESIGN STRUCTURES . .
87
3.2.11 BINDING OF OTHER STEROIDS IN THE DESIGNED POCKET 91
3.2.12 CRYSTAL STRUCTURE OF DBG-2 IN THE LOW-AFFINITY CLEAVED STATE ....
93
3.2.13 COMPARISON OF NATIVE AND CLEAVED ACT DESIGNS AND THE TRIGGERED
STEROID RELEASE 95
4 DISCUSSION 105
4.1 CBG 105
4.1.1 RCL CLEAVAGE AND S-TO-R TRANSITION AS EMERGENCY MECHANISM FOR
STEROID RELEASE 105
4.1.2 RAT CBG DEVIATES FROM THE CANONICAL SERPIN MECHANISM 106
4.1.3 CBG SEQUENCE CONSERVATION AND FUNCTION 113
4.1.4 THE FLEXIBLE SERPIN FOLD AND GENETIC VARIANTS IN CBG 115
4.1.5 POSSIBLE INFLUENCES OF POSTTRANSLATIONAL MODIFICATIONS ON DYNAMICS
WITHIN THE SERPIN FOLD 117
4.2 DESIGN OF AN ACT-BASED MODEL SYSTEM 119
4.2.1 POSSIBILITIES AND ACHIEVEMENTS IN USING MUMBO TO GUIDE DESIGN .
119
4.2.2 FEATURES OF STEROID BINDING IN ACT DESIGNS 120
4.2.3 CONFORMATIONAL CHANGE IN ACT DESIGNS 123
4.3 ALLOSTERIC REGULATION OF LIGAND RELEASE IN SERPINS 123
BIBLIOGRAPHY 127
5 APPENDIX 145
IV
|
| any_adam_object | 1 |
| author | Gardill, Bernd R. |
| author_facet | Gardill, Bernd R. |
| author_role | aut |
| author_sort | Gardill, Bernd R. |
| author_variant | b r g br brg |
| building | Verbundindex |
| bvnumber | BV041711533 |
| ctrlnum | (OCoLC)876305881 (DE-599)DNB1045597201 |
| dewey-full | 571.74 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.74 |
| dewey-search | 571.74 |
| dewey-sort | 3571.74 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| id | DE-604.BV041711533 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T01:03:30Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-027158712 |
| oclc_num | 876305881 |
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| owner | DE-188 DE-29T DE-29 DE-12 DE-355 DE-BY-UBR |
| owner_facet | DE-188 DE-29T DE-29 DE-12 DE-355 DE-BY-UBR |
| physical | VI, 156 S. Ill., graph. Darst. |
| publishDate | 2013 |
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| spelling | Gardill, Bernd R. Verfasser aut Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG vorgelegt von Bernd Rainer Gardill Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG 2013 VI, 156 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Erlangen-Nürnberg, Univ., Diss., 2013 Allosterische Kontrolle (DE-588)4141976-5 gnd rswk-swf Corticosteroid-bindendes Globulin (DE-588)4148330-3 gnd rswk-swf Serpine (DE-588)4285093-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Corticosteroid-bindendes Globulin (DE-588)4148330-3 s Serpine (DE-588)4285093-9 s Allosterische Kontrolle (DE-588)4141976-5 s DE-604 application/pdf http://d-nb.info/1045597201/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027158712&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gardill, Bernd R. Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG Allosterische Kontrolle (DE-588)4141976-5 gnd Corticosteroid-bindendes Globulin (DE-588)4148330-3 gnd Serpine (DE-588)4285093-9 gnd |
| subject_GND | (DE-588)4141976-5 (DE-588)4148330-3 (DE-588)4285093-9 (DE-588)4113937-9 |
| title | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG |
| title_alt | Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG |
| title_auth | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG |
| title_exact_search | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG |
| title_full | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG vorgelegt von Bernd Rainer Gardill |
| title_fullStr | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG vorgelegt von Bernd Rainer Gardill |
| title_full_unstemmed | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG vorgelegt von Bernd Rainer Gardill |
| title_short | Unravelling the allosteric mechanism that links hormone release in CBG to the serpin-specific stressed to relaxed transition |
| title_sort | unravelling the allosteric mechanism that links hormone release in cbg to the serpin specific stressed to relaxed transition erforschung des allosterischen kopplungsmechanismus zwischen serpin spezifischer umlagerung und hormonfreisetzung in cbg |
| title_sub | = Erforschung des allosterischen Kopplungsmechanismus zwischen Serpin-spezifischer Umlagerung und Hormonfreisetzung in CBG |
| topic | Allosterische Kontrolle (DE-588)4141976-5 gnd Corticosteroid-bindendes Globulin (DE-588)4148330-3 gnd Serpine (DE-588)4285093-9 gnd |
| topic_facet | Allosterische Kontrolle Corticosteroid-bindendes Globulin Serpine Hochschulschrift |
| url | http://d-nb.info/1045597201/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027158712&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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