Functional and structural characterization of the virulence factor IpaB from Shigella flexneri:
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Berlin
Mensch-und-Buch-Verl.
2009
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | IX, 97 S. Ill., graph. Darst. |
| ISBN: | 9783866647398 |
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MARC
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| 245 | 1 | 0 | |a Functional and structural characterization of the virulence factor IpaB from Shigella flexneri |c vorgelegt von Lidija Senerovic |
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| adam_text | TABLE OF CONTENTS IM_ TABLE OF CONTENTS 1 INTRODUCTION 1 1.1 SHIGELLA-
THE ETIOLOGICAL AGENT OF DYSENTERY 1 1.2 CELLULAR PATHOGENESIS OF
SHIGELLOSIS 1 1.3. MOLECULAR BASIS OF S. FLEXNERI VIRULENCE 3 1.3.1 TYPE
III SECRETION SYSTEM 3 1.3.2 SHIGELLA VIRULENCE FACTORS 4 1.3.2.1
INVASION PLASMID ANTIGEN B (IPAB) 4 1.3.2.2 OTHER VIRULENCE FACTORS
REQUIRED FOR S. FLEXNERI PATHOGENESIS 6 1.4 CELL DEATH DURING INFECTION
7 1.4.1 PROGRAMMED CELL DEATH 8 1.4.2 CASPASES 10 1.4.2.1 CASPASE-1 10
1.4.2.2 NLR AND THE INFLAMASOMES 11 1.4.2.3 IL-1P AND IL-18 13 1.4.3
CASPASE-INDEPENDENT CELL DEATH 14 1.5 AIMS OF THE STUDY 15 2 RESULTS 17
2.1 FUNCTIONAL STUDIES ON IPAB 17 2.1.1 IPAB PURIFICATION 17 2.1.2
PURIFIED IPAB HAS A STABLE CORE 19 2.1.3 IPAB FORMS OLIGOMERS IN
SOLUTION 21 2.1.4 IPAB OLIGOMERS PERMEABILIZE MEMBRANES 22 2.1.5 IPAB
FORMS CHANNELS SELECTIVE FOR MONOVALENT CATIONS 25 2.1.6 IPAB KILLS
PHAGOCYTES BUT NOT EPITHELIAL CELLS 27 2.1.7 IPAB CYTOTOXICITY REQUIRES
INTERNALIZATION 29 2.1.8 IPAB INDUCES LEAKAGE OF LYSOSOMES 31 2.1.9 IPAB
ACTIVATES CASPASE-1 THROUGH THE IPAF INFLAMMASOME 33 2.1.10 IPAC DOES
NOT CONTRIBUTE TO IPAB FUNCTION 35 2.2 STRUCTURAL ANALYSIS OF IPAB 37
2.2. BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/1000281930
DIGITALISIERT DURCH TABLE OF CONTENTS 3 DISCUSSION 42 3.1 PURIFICATION
OF IPAB USING DETERGENTS 44 3.2 IPAB FORMS ION CHANNELS 45 3.3 IPAB ION
CHANNELS DAMAGE ENDOLYSOSOMES 47 3.4 IPAB KILLS PHAGOCYTES 48 3.5 IPAB
AS PART OF A TRANSLOCON 50 3.6 PROPOSED MODEL FOR IPAB- INDUCED
MACROPHAGE DEATH 51 3.7 STRATEGIES TO CRYSTALLIZE IPAB 52 3.8 STRUCTURE
OF LPAB95-290 54 4 MATERIALS AND METHODS 57 4.1 MATERIALS 57 4.1.1
STRAINS AND CELL LINES 57 4.1.2 PLASMIDS 58 4.1.3 DNA OLIGOS FOR CLONING
59 4.1.4 ANTIBODIES 60 4.1.5 CULTURE MEDIA AND ANTIBIOTICS 60 4.2
METHODS 61 4.2.1 MOLECULAR BIOLOGY METHODS 61 4.2.1.1 CONSTRUCTION OF
PLASMIDS 61 4.2.1.2 AGAROSE GEL ELECTROPHORESIS AND GEL EXTRACTION 62
4.2.1.3 PREPARATION OF ELECTROCOMPETENT E. COLI 62 4.2.1.4
TRANSFORMATION 62 4.2.1.5 ISOLATION OF PLASMID DNA 62 4.2.1.6 LIGATION
63 4.2.2 TISSUE CULTURE 63 4.2.2.1 MOUSE MACROPHAGES 63 4.2.2.2 HUMAN
BLOOD MONOCYTES AND MACROPHAGES 63 4.2.3. CELLULAR MICROBIOLOGY METHODS
64 4.2.3.1 BACTERIAL INFECTIONS 64 4.2.3.2 CASPASE-1 ACTIVITY ASSAY 64
4.2.3.3 LDH RELEASE ASSAY 64 4.2.3.4 SYTOXGREEN BASED CYTOTOXICITY ASSAY
65 4.2.3.5 HEMOLYSIS 65 4.2.3.6 FLOW CYTOMETRY 65 4.2.3.7
IMMUNOFLUORESCENCE 65 4.2.3.8 IMMUNOELECTRON MICROSCOPY 66 4.2. TABLE OF
CONTENTS 4.2.5.1 PROTEIN EXPRESSION 67 4.2.5.2 PROTEIN PURIFICATION 68
4.2.5.3 SODIUM DODECYLSULFATE POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-
PAGE) 69 4.2.5.4 NATIVE-PAGE 69 4.2.5.5 STAINING OF PAGE GELS 69 4.2.5.6
DETERMINATION OF PROTEIN CONCENTRATION 70 4.2.5.7 LIMITED PROTEOLYSIS 70
4.2.5.8 WESTERN TRANSFER 70 4.2.5.9 CHEMICAL CROSSLINKING 71 4.2.5.10
MASS SPECTROMETRY 71 4.2.5.11 LIPOSOME LEAKAGE ASSAY 71 4.2.5.12
ENZYME-LINKED IMMUNOSORBENT ASSAY 72 4.2.6 SPECTROSCOPIC METHODS 73
4.2.6.1 CRYSTALLIZATION 73 4.2.6.2 CRYSTAL FREEZING 73 4.2.6.3 DATA
COLLECTION AND PROCESSING 73 4.2.6.4 CRYSTAL STRUCTURE DETERMINATION 73
4.2.6.5 MULTI-ANGLE LASER LIGHT SCATTERING (MALLS) 74 5 REFERENCES 75 6
APPENDIX 93
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| any_adam_object | 1 |
| author | Senerovic, Lidija 1977- |
| author_GND | (DE-588)140582541 |
| author_facet | Senerovic, Lidija 1977- |
| author_role | aut |
| author_sort | Senerovic, Lidija 1977- |
| author_variant | l s ls |
| building | Verbundindex |
| bvnumber | BV036477608 |
| classification_rvk | XD 5000 |
| ctrlnum | (OCoLC)607572627 (DE-599)BVBBV036477608 |
| dewey-full | 616.07995 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 616 - Diseases |
| dewey-raw | 616.07995 |
| dewey-search | 616.07995 |
| dewey-sort | 3616.07995 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| indexdate | 2024-07-09T22:40:19Z |
| institution | BVB |
| isbn | 9783866647398 |
| language | English |
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| physical | IX, 97 S. Ill., graph. Darst. |
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| publisher | Mensch-und-Buch-Verl. |
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| spelling | Senerovic, Lidija 1977- Verfasser (DE-588)140582541 aut Functional and structural characterization of the virulence factor IpaB from Shigella flexneri vorgelegt von Lidija Senerovic Berlin Mensch-und-Buch-Verl. 2009 IX, 97 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: Berlin, Humboldt-Univ., Dis., 2009 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020349183&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Senerovic, Lidija 1977- Functional and structural characterization of the virulence factor IpaB from Shigella flexneri |
| subject_GND | (DE-588)4113937-9 |
| title | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri |
| title_auth | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri |
| title_exact_search | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri |
| title_full | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri vorgelegt von Lidija Senerovic |
| title_fullStr | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri vorgelegt von Lidija Senerovic |
| title_full_unstemmed | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri vorgelegt von Lidija Senerovic |
| title_short | Functional and structural characterization of the virulence factor IpaB from Shigella flexneri |
| title_sort | functional and structural characterization of the virulence factor ipab from shigella flexneri |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020349183&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT seneroviclidija functionalandstructuralcharacterizationofthevirulencefactoripabfromshigellaflexneri |