Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants: characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Online Access: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20121114-1108313-1-1 Inhaltsverzeichnis |
| Physical Description: | 78 Bl. Ill., graph. Darst. |
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| 100 | 1 | |a Brunie, Leonora |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants |b characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |c Leonora Petrova Brunie |
| 264 | 1 | |c 2012 | |
| 300 | |a 78 Bl. |b Ill., graph. Darst. | ||
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| 502 | |a München, Techn. Univ., Diss., 2012 | ||
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Record in the Search Index
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| adam_text | IMAGE 1
TABLE O F CONTENTS
TABLE OF CONTENTS
1 INTRODUCTION 1
1.1 THE INTEGRIN FAMILY 1
1.2 INTEGRIN-MEDIATED SIGNALING 3
1.3 INTEGRIN AVP3 4
1.3.1 STRUCTURE O F INTEGRIN AVP3 6
1.3.2 FUNCTIONS O F INTEGRIN AVP3 IN ANGIOGENESIS AND CANCER PROGRESSION
9
1.3.3 INTEGRIN AVP3 CROSSTALK WITH THE EPIDERMAL GROWTH FACTOR RECEPTOR
11
1.4 CONFORMATIONAL CHANGES DURING INTEGRIN ACTIVATION 12
1.5 AIM O F THE WORK 15
2 MATERIALS AND METHODS 16
2.1 MATERIALS 16
2.1.1 CELL LINE 16
2.1.2 BACTERIAL STRAIN 16
2.1.3 CELL CULTURE REAGENTS 16
2.1.4 GENERAL CHEMICALS 16
2.1.5 GENERAL SOLUTIONS AND BUFFERS 17
2.1.6 EXPRESSION PLASMID 19
2.1.7 ANTIBIOTICS 20
2.1.8 ENZYMES 20
2.1.9 KITS 20
2.1.10 PRIMERS FOR IN VITRO SITE-DIRECTED MUTAGENESIS 21
2.1.11 ANTIBODIES 22
2.1.12 INSTRUMENTS 22
2.2 METHODS 23
2.2.1 CELL CULTURE 23
2.2.2 IN VITRO SITE-DIRECTED MUTAGENESIS 24
2.2.3 AMPLIFICATION, PURIFICATION, AND ANALYSIS O F PLASMID DNA 26
2.2.4 CELL PROLIFERATION ASSAYS 28
2.2.5 FLOW CYTOFLUOROMETRY 29
3 RESULTS 31
3.1 GENERATION O F INTEGRIN AVP3 MUTANTS 31
3.1.1 GENERATION O F INTEGRIN AVP3 MUTANTS BY IN VITRO SITE-DIRECTED
MUTAGENESIS 33
HTTP://D-NB.INFO/1033367117
IMAGE 2
TABLE O F CONTENTS
3.1.2 GENERATION O F MUTANTS O F THE INTEGRIN AVP3 SALT BRIDGE 34
3.1.3 GENERATION O F MUTANTS O F THE INTEGRIN P3 TRANSMEMBRANE DOMAIN 37
3.1.3.1 EXCHANGE O F THE INTEGRIN 03 TRANSMEMBRANE DOMAIN BY THAT O F
GLYCOPHORIN A 37
3.1.3.2 GENERATION O F A POINT MUTATED GXXXG MOTIF O F THE GLYCOPHORIN A
TRANSMEMBRANE DOMAIN TO GXXXL 48
3.2 ESTABLISHMENT O F HUMAN OVARIAN CANCER CELL TRANSFECTANTS EXPRESSING
EITHER WILD TYPE OR DISTINCT MUTANT FORMS O F INTEGRIN AVP3 51
3.2.1 IMMUNOCYTOCHEMICAL DETECTION O F THE CONTENT O F INTEGRIN AVP3 AND
ITS VARIANTS IN TRANSFECTED HUMAN OVARIAN CANCER CELLS 52
3.3 INTEGRIN AVP3-MEDIATED CELL PROLIFERATION AS A FUNCTION O F ITS
TRANSMEMBRANE
DOMAIN SEQUENCE 52
3.4 COOPERATION BETWEEN INTEGRIN AVP3 AND THE EPIDERMAL GROWTH FACTOR
RECEPTOR 56
3.4.1 EPIDERMAL GROWTH FACTOR RECEPTOR EXPRESSION AS A FUNCTION O F THE
INTEGRIN AVP3
TRANSMEMBRANE DOMAIN SEQUENCE 56
3.4.2 EFFECT O F EPIDERMAL GROWTH FACTOR ON INTEGRIN AVP3-DEPENDANT
HUMAN OVARIAN
CANCER CELL PROLIFERATION AS A FUNCTION O F INTEGRIN TRANSMEMBRANE
DOMAIN
CONFORMATION 58
4 DISCUSSION 59
4.1 IN VITRO SITE-DIRECTED MUTAGENESIS O F INTEGRIN AVP3 59
4.2 IMPACT O F INTEGRIN CYTOPLASMIC SALT BRIDGE FORMATION ON INTEGRIN
AVP3
ACTIVATION 61
4.3 IMPACT O F THE INTEGRIN AVP3 TRANSMEMBRANE DOMAIN ON HUMAN OVARIAN
CANCER CELL PROLIFERATION 64
4.4 EPIDERMAL GROWTH FACTOR RECEPTOR EXPRESSION AS A FUNCTION OF
THE INTEGRIN AVP3 TRANSMEMBRANE DOMAIN CONFORMATION 66
4.5 SUMMARY 68
5 LIST OF FIGURES 70
6 REFERENCES 72
|
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| spelling | Brunie, Leonora Verfasser aut Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation Leonora Petrova Brunie 2012 78 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2012 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20121114-1108313-1-1 http://mediatum.ub.tum.de/node?id=1108313 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20121114-1108313-1-1 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025767765&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Brunie, Leonora Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |
| subject_GND | (DE-588)4113937-9 |
| title | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |
| title_auth | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |
| title_exact_search | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |
| title_full | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation Leonora Petrova Brunie |
| title_fullStr | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation Leonora Petrova Brunie |
| title_full_unstemmed | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation Leonora Petrova Brunie |
| title_short | Generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants |
| title_sort | generation of integrin alphavbeta3 cytoplasmic and transmembrane domain mutants characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |
| title_sub | characterisation of the impact of integrin conformation on human ovarian cancer cell proliferation |
| topic_facet | Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1108313 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20121114-1108313-1-1 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025767765&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT brunieleonora generationofintegrinalphavbeta3cytoplasmicandtransmembranedomainmutantscharacterisationoftheimpactofintegrinconformationonhumanovariancancercellproliferation |