Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
|
| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120920-1108725-0-3 Inhaltsverzeichnis |
| Beschreibung: | 113 Blatt Ill., graph. Darst. |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV041064400 | ||
| 003 | DE-604 | ||
| 005 | 20130705 | ||
| 007 | t | ||
| 008 | 130604s2012 ad|| m||| 00||| eng d | ||
| 035 | |a (OCoLC)854721702 | ||
| 035 | |a (DE-599)BVBBV041064400 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-91 |a DE-12 | ||
| 082 | 0 | |a 610 | |
| 084 | |a CHE 890f |2 stub | ||
| 084 | |a CHE 832f |2 stub | ||
| 100 | 1 | |a Schneider, Elisabeth Veronika |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex |c Elisabeth V. Schneider |
| 264 | 1 | |c 2012 | |
| 300 | |a 113 Blatt |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Techn. Univ., Diss., 2012 | ||
| 546 | |a Mit einer Zssfassung in engl. Sprache | ||
| 650 | 0 | 7 | |a RNS-Polymerase II |0 (DE-588)4318423-6 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a RNS-Polymerase II |0 (DE-588)4318423-6 |D s |
| 689 | 0 | |5 DE-604 | |
| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:91-diss-20120920-1108725-0-3 |
| 856 | 4 | 1 | |u http://mediatum.ub.tum.de/node?id=1108725 |x Verlag |z kostenfrei |3 Volltext |
| 856 | 4 | |u https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120920-1108725-0-3 |x Resolving-System | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026041447&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-026041447 | ||
Datensatz im Suchindex
| _version_ | 1804150430267080704 |
|---|---|
| adam_text | IMAGE 1
INDEX
INDEX
1. INTRODUCTION 1
1.1 CANCEROGENESIS 1
1.2 THE REGULATION OF THE CELL CYCLE AND THE CELL CYCLE CDKS 1
1.3 THE TRANSCRIPTIONAL PROCESS AND THE INVOLVEMENT OF THE CTD KINASES 3
1.4 THE STRUCTURAL ORGANIZATION OF THE CDK/CYCLIN FAMILY AND ITS
IMPLICATION ON THE CDK. KINASE FUNCTION 5
1.5 CDK8/CYCC IS A PROMINENT MEMBER OF THE CDK FAMILY DUE TO ITS ROLE IN
PHYSIOLOGY AND DISEASE 9
1.6 DRUG-ABILITY OF CDK8/CYCC: TYPE I AND TYPE II INHIBITORS 10
1.7 AIMS OF THIS THESIS 13
2. MATERIAL AND METHODS 14
2.1 MOLECULAR CLONING 14
2.1.1 POLYMERASE CHAIN REACTION (PCR) BASED DNA AMPLIFICATION AND
SUB-CLONING 14 2.1.2 BACTERIAL TRANSFORMATION 15
2.2 SITE-DIRECTED MUTAGENESIS 16
2.3 BACTERIAL EXPRESSION 17
2.4 INSECT CELL CULTURE AND HETEROLOGOUS PROTEIN EXPRESSION 17
2.5 ANALYSIS OF TESTEXPRESSIONS 18
2.6 SODIUM DODECYL SULFATE - POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE) ANALYSIS AND PROTEIN STAINING 18
2.7 WESTERN BLOT ANALYSIS 19
2.8 DETERMINATION OF PROTEIN AND NUCLEIC ACID CONCENTRATION 19
2.9 HIS-TAG/GST-TAG PURIFICATION 19
2.10 SIZE EXCLUSION CHROMATOGRAPHY (SEC) 20
2.11 PROTEINS CONCENTRATION STEPS 20
2.12 PURIFICATION PROTOCOLS 20
2.13 CRYSTALLIZATION, X-RAY DATA COLLECTION, PROCESSING AND REFINEMENT
21
2.14 CDK8/CYCC KD MEASUREMENT 23
2.15 THE PROTEROS REPORTER DISPLACEMENT ASSAY 24
2.16 CHEMICALS 26
3. ESTABLISHMENT OF PROTEIN PRODUCTION, CRYSTALLIZATION AND FIRST
STRUCTURE DETERMINATION OF THE HUMAN CDK8/CYCC/SORAFENIB COMPLEX 27
3.1 TESTING THE DIFFERENT CYCC-CONSTRUCTS IN E. COLI SHOWS THAT IT IS
NOT A SUITABLE EXPRESSION SYSTEM FOR CYCC BUT REVEALS THE MOST
APPROPRIATE TAG-FUSION CONSTRUCT 27 3.2 PROTEIN EXPRESSION OF CDK8 AND
CYCC IS ONLY POSSIBLE SEPARATELY IN SF9 CELLS 32 3.3 ESTABLISHMENT OF
THE CDK8/CYCC COMPLEX FORMATION IN-VITRO AND ITS PURIFICATION 36
3.4 CRYSTALLIZATION AND REFINEMENT OF CRYSTALS OF THE
CDK8/CYCC/SORAFENIB COMPLEX 38 3.5 DETERMINATION OF THE CRYSTAL
STRUCTURE OF THE CDK8/CYCC/SORAFENIB COMPLEX 40
4. THE FIRST STRUCTURE OF CDK8/CYCC WITHIN THE CDK8-DMG-OUT CONFORMATION
IMPLICATES SPECIFICITY WITHIN THE CDK/CYCLIN FAMILY AND REVEALS
INTERACTION WITH A DEEP POCKET BINDER 44
4.1 THE OVERALL STRUCTURE OF CDK8/CYCC 44
4.2 CRYSTAL STRUCTURE OF HUMAN CYCC 46
4.3 CRYSTAL STRUCTURE OF HUMAN CDK8 49
HTTP://D-NB.INFO/1038308925
IMAGE 2
INDEX
4.4 THE CONSERVED CORE OF THE CDK/CYCLIN INTERFACE OF CDK8/CYCC:
COMPARISON WITH CDK9/CYCTL 52
4.5 THE ADAPTABLE H N HELIX OF CYCC IS CONTACTING CDK8 54
4.6 CDK8 CONTAINS AN ADDITIONAL N-TERMINAL CYCC RECOGNITION HELIX 55
4.7 SORAFENIB BINDS TO THE HINGE REGION IN CDK.8 INDUCING DMG-OUT
CONFORMATION 58
5. THE DISCOVERY OF SLOWNESS IN TERMS OF COMPOUND BINDING KINETICS: A
STRUCTURE-KINETIC-RELATIONSHIP STUDY ON CDK8/CYCC 61
5.1 THE PRIMARY SCREEN AND THE DISCOVERY OF SLOWNESS 61
5.2 COMBINING STRUCTURAL WITH KINETIC DATA REVEALS THAT THE FLIP OF THE
CDK8 DMG-MOTIF DOES NOT INFLUENCE THE KINETIC BINDING BEHAVIOR OF THE
COMPOUNDS OF THE SKR SE RIES 67 5.3 THE X-RAY STRUCTURES OF CDK8/CYCC
WITH SLOW AND FAST BINDING COMPOUNDS INDICATE THAT THE MAIN
CONTRIBUTIONS TO RESIDENCE TIME ARE CONSERVED
HYDROGEN-BONDING AND HYDROPHOBIC CONTACTS WITHIN THE CDK.8 FRONT POCKET
75 5.4 DISCUSSION 81
6. CDK8/CYCC IN THE DMG-IN KINASE CONFORMATION VERSUS THE CDK8/CYCC
DMG-OUT KINASE CONFORMATION: THE TWO FACES OF THE CDK8/CYCC COMPLEX 85
6.1 BINDING ASSAY TO SELECT POTENTIAL CDK8/CYCC INHIBITORS THAT TARGET
THE CDK8 DMG-IN KINASE CONFORMATION 85
6.2 STRUCTURAL DATA ON COMPOUND 12 IN THE ACTIVE SITE CDK8/CYCC SITE 88
6.3 ARRANGEMENTS OF CDK8/CYCC RELATED TO THE DMG-IN CONFORMATION 90
6.4 THE ACTIVE SITE OF CDK8/CYCC IN ABSENCE OF AN INHIBITOR 93
6.5 THE CRUCIAL INFLUENCE OF CYCC BINDING ON THE CDK.8 CONFORMATION
ALLOWS FOR SELECTIVE INHIBITION OF CDK8/CYCC AND DISCRIMINATION AGAINST
THE CDK8 CONFORMATION IN ABSENCE OF THE CYCLIN 96
6.6 COMPARISON OF THE CYCC DEPENDENCE OF FURTHER SKR COMPOUNDS 99
7. REFERENCES 103
CURRICULUM VITAE 113
|
| any_adam_object | 1 |
| author | Schneider, Elisabeth Veronika |
| author_facet | Schneider, Elisabeth Veronika |
| author_role | aut |
| author_sort | Schneider, Elisabeth Veronika |
| author_variant | e v s ev evs |
| building | Verbundindex |
| bvnumber | BV041064400 |
| classification_tum | CHE 890f CHE 832f |
| collection | ebook |
| ctrlnum | (OCoLC)854721702 (DE-599)BVBBV041064400 |
| dewey-full | 610 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
| dewey-search | 610 |
| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Chemie Medizin |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01727nam a2200409 c 4500</leader><controlfield tag="001">BV041064400</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20130705 </controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">130604s2012 ad|| m||| 00||| eng d</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)854721702</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BVBBV041064400</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-91</subfield><subfield code="a">DE-12</subfield></datafield><datafield tag="082" ind1="0" ind2=" "><subfield code="a">610</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">CHE 890f</subfield><subfield code="2">stub</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">CHE 832f</subfield><subfield code="2">stub</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Schneider, Elisabeth Veronika</subfield><subfield code="e">Verfasser</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex</subfield><subfield code="c">Elisabeth V. Schneider</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2012</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">113 Blatt</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="a">München, Techn. Univ., Diss., 2012</subfield></datafield><datafield tag="546" ind1=" " ind2=" "><subfield code="a">Mit einer Zssfassung in engl. Sprache</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">RNS-Polymerase II</subfield><subfield code="0">(DE-588)4318423-6</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="689" ind1="0" ind2="0"><subfield code="a">RNS-Polymerase II</subfield><subfield code="0">(DE-588)4318423-6</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2=" "><subfield code="5">DE-604</subfield></datafield><datafield tag="776" ind1="0" ind2="8"><subfield code="i">Erscheint auch als</subfield><subfield code="n">Online-Ausgabe</subfield><subfield code="o">urn:nbn:de:bvb:91-diss-20120920-1108725-0-3</subfield></datafield><datafield tag="856" ind1="4" ind2="1"><subfield code="u">http://mediatum.ub.tum.de/node?id=1108725</subfield><subfield code="x">Verlag</subfield><subfield code="z">kostenfrei</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="856" ind1="4" ind2=" "><subfield code="u">https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120920-1108725-0-3</subfield><subfield code="x">Resolving-System</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026041447&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ebook</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-026041447</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV041064400 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T00:38:48Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-026041447 |
| oclc_num | 854721702 |
| open_access_boolean | 1 |
| owner | DE-91 DE-BY-TUM DE-12 |
| owner_facet | DE-91 DE-BY-TUM DE-12 |
| physical | 113 Blatt Ill., graph. Darst. |
| psigel | ebook |
| publishDate | 2012 |
| publishDateSearch | 2012 |
| publishDateSort | 2012 |
| record_format | marc |
| spelling | Schneider, Elisabeth Veronika Verfasser aut Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex Elisabeth V. Schneider 2012 113 Blatt Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2012 Mit einer Zssfassung in engl. Sprache RNS-Polymerase II (DE-588)4318423-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content RNS-Polymerase II (DE-588)4318423-6 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20120920-1108725-0-3 http://mediatum.ub.tum.de/node?id=1108725 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120920-1108725-0-3 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026041447&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schneider, Elisabeth Veronika Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex RNS-Polymerase II (DE-588)4318423-6 gnd |
| subject_GND | (DE-588)4318423-6 (DE-588)4113937-9 |
| title | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex |
| title_auth | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex |
| title_exact_search | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex |
| title_full | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex Elisabeth V. Schneider |
| title_fullStr | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex Elisabeth V. Schneider |
| title_full_unstemmed | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex Elisabeth V. Schneider |
| title_short | Structural and functional characterization of human protein kinases exemplified on the human cyclin-dependent kinase 8- cyclin C complex |
| title_sort | structural and functional characterization of human protein kinases exemplified on the human cyclin dependent kinase 8 cyclin c complex |
| topic | RNS-Polymerase II (DE-588)4318423-6 gnd |
| topic_facet | RNS-Polymerase II Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1108725 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120920-1108725-0-3 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026041447&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schneiderelisabethveronika structuralandfunctionalcharacterizationofhumanproteinkinasesexemplifiedonthehumancyclindependentkinase8cyclinccomplex |