Role of the novel protein tyrosine phosphatase AUM for cell adhesion:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
2011
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| Subjects: | |
| Online Access: | Volltext Inhaltsverzeichnis |
| Item Description: | Online erschienen: 2013 |
| Physical Description: | VII, 112 S. Ill., graph. Darst. |
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MARC
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| 100 | 1 | |a Saxena, Ambrish |d 1979- |e Verfasser |0 (DE-588)1044452277 |4 aut | |
| 245 | 1 | 0 | |a Role of the novel protein tyrosine phosphatase AUM for cell adhesion |c von Ambrish Saxena |
| 246 | 1 | 3 | |a Die Rolle des neuen Proteins "Tyrosin phosphatase" AUM für Zell-Adhäsion |
| 264 | 1 | |c 2011 | |
| 300 | |a VII, 112 S. |b Ill., graph. Darst. | ||
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Record in the Search Index
| _version_ | 1804151553513226240 |
|---|---|
| adam_text | CONTENTS
I
CONTENTS
1 INTRODUCTION
: :
1
1.1 CLASSIFICATION OF PHOSPHATASES 2
HAD-SUPERFAMILY OF PHOSPHATASES
3
CHRONOPHIN
3
1.2 IDENTIFICATION AND CHARACTERIZATION OF NOVEL HAD-PHOSPHATASE AUM
3
1.3
REGULATION OF CELL ADHESION BY PHOSPHATASES AND KINASES
6
1.4
CELL ADHESION ON EXTRACELLULAR MATRIX
8
FIBRONECTIN 8
1.5 INTEGRINS 9
INTEGRIN ACTIVATION 10
BIDIRECTIONAL SIGNALING THROUGH INTEGRINS 11
INTEGRIN BINDING PROTEINS 14
1.6
FOCAL ADHESIONS
15
1.7
ROLE OF ACTIN DYNAMICS FOR CELL ADHESION ON EXTRACELLULAR MATRIX
17
1.8 REGULATION OF ADHESION DYNAMICS BY RHOGTPASES 18
DOWNSTREAM EFFECTORS OF RHOA 19
REGULATION OF RHOA 20
ROLE OF INTEGRINS IN RHOA ACTIVATION 21
2 AIM OF THE STUDY 22
3 MATERIALS 23
3.1 LIST OF MANUFACTURERS AND DISTRIBUTORS 23
3.2 CHEMICALS 24
33 NUCLEOTIDES, NUCLEIC ACIDS AND PRIMERS 25
3.4 PLASMIDS 25
3.5 ANTIBODIES 25
3.6 BACTERIAL STRAIN AND CELL LINES 26
3.7 TISSUE CULTURE REAGENTS AND MATERIALS 27
3.8
OTHER
MATERIALS
27
3.9 COMMERCIAL KITS 28
3.10 SOFTWARE AND
DATABASES 28
3.11 RNA INTERFERENCE TOOLS 28
3.12 SOLUTIONS & BUFFERS 29
4 METHODS 34
4.1 CELL BIOLOGY METHODS 34
CELL LINES 34
TRANSIENT TRANSFECHON 34
IMMUNOCYTOCHEMISTRY 35
CONFOCAL LASER-SCANNING MICROSCOPY 36
RNA INTERFERENCE 36
VALIDATION OF THE RNAI TOOLS 37
4.2 CELL BIOLOGICAL ASSAYS 39
CELL SPREADING ON FIBRONECTIN 39
CELL AREA DETERMINATION OF FIXED CELLS 39
CELL- ECM ADHESION ASSAY 39
43
FLOW CYTOMETRY 41
HTTP://D-NB.INFO/1045120200
CONTENTS
II
4.4 MOLECULAR BIOLOGY METHODS 42
PLASMID PREPARATION 42
QUANTIFICATION OF NUCLEIC ACIDS BY PHOTOMETRIC MEASUREMENT 43
RESTRICTION DIGESTS OF PLASMID DNA 43
DNA GEL ELECTROPHORESIS 43
SITE-DIRECTED MUTAGENESIS 43
4.5 PROTEIN BIOCHEMICAL METHODS 44
SAMPLE PREPARATION FOR
IMMUNOBLOT ANALYSIS 44
ESTIMATION OF PROTEIN CONCENTRATION 44
SDS-POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-PAGE) 44
IMMUNOBLOT ANALYSIS 45
REPROBING OF NITROCELLULOSE MEMBRANES 46
4.6 RHOA/RACI ACTIVITY
ASSAY 46
ACTIVE RHO/RAC PULL-DOWN
ASSAY 47
ACTIVE RHOA/RAC
1 G-LISA ASSAY 47
5 RESULTS * * ** * * 48
5.1 AUM KNOCKDOWN BY RNA INTERFERENCE 48
STABLE AUM DEPLETION 48
TRANSIENT AUM DEPLETION 49
5.2 AUM IS INVOLVED IN EARLY CELL SPREADING 50
5.3 AUM REGULATES EARLY CELL-ECM ADHESION 52
GC-1 CELLS PREFERENTIALLY ADHERE TO FN 52
EFFECT OF AUM ON CELL-FN ADHESION 52
5.4 ACUTE AUM DEPLETION
ALSO LEADS TO INCREASED CELL ADHESION ON
FN 55
5.5 RE-EXPRESSION OF AUM NORMALIZES CELL ADHESION 56
5.6 ACCELERATED ADHESION DUE TO AUM-DEPLETION IS INTEGRIN DEPENDENT 60
5.7 CELL SURFACE EXPRESSION OF INTEGRINS 61
5.8 AUM-DEPLETED CELLS HAVE MORE ACTIVE P1-INTEGRIN ON
CELL SURFACE 64
P1 -INTEGRIN ACTIVATION IS
FN-DEPENDENT 65
5.9 AUM INFLUENCES ACTIN DYNAMICS DURING ADHESION AND SPREADING 67
5.10 AUM IS A NEGATIVE REGULATOR OF RHOA 68
INCREASED ADHESION DUE TO AUM-DEPLETION IS
RHOA/ROCK DEPENDENT 70
5.11 AUM-DEPLETION INDUCES FASTER FOCAL
ADHESION MATURATION 73
5.12 AUM-DEPLETED CELLS HAVE MORE PHOSPHOR-TYROSINE CONTENT 75
6 DISCUSSION 77
6.1 EFFECT OF AUM ON CELL ADHESION
AND SPREADING 78
6.2 EFFECT OF AUM DEPLETION ON CELL
ADHESION CAN BE RESCUED 79
6.3 INTEGRINS ARE INVOLVED IN AUM MEDIATED CELL ADHESION 80
6.4 EFFECT OF AUM ON INTEGRIN
ACTIVATION 81
6.5 AUM MODULATES ACTIN DYNAMICS MEDIATED BY ITS EFFECT ON RHOA 82
6.6 ROLE OF AUM IN FOCAL
ADHESION FORMATION
AND MATURATION 83
6.7 POSSIBLE MODEL FOR ROLE OF
AUM IN CELL ADHESION AND SPREADING 84
7 SUMMARY 86
8 ZUSAMMENFASSUNG 88
9 REFERENCES 90
CONTENTS JLL
10 CURRICULUM VITAE 107
U ERKLARUNG 109
12 ACKNOWLEDGEMENTS 110
|
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| language | English |
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| spelling | Saxena, Ambrish 1979- Verfasser (DE-588)1044452277 aut Role of the novel protein tyrosine phosphatase AUM for cell adhesion von Ambrish Saxena Die Rolle des neuen Proteins "Tyrosin phosphatase" AUM für Zell-Adhäsion 2011 VII, 112 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Online erschienen: 2013 Würzburg, Univ., Diss., 2011 Zelladhäsion (DE-588)4314136-5 gnd rswk-swf Proteintyrosinphosphatase (DE-588)4311372-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Zelladhäsion (DE-588)4314136-5 s Proteintyrosinphosphatase (DE-588)4311372-2 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-65503 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-65503 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026883678&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Saxena, Ambrish 1979- Role of the novel protein tyrosine phosphatase AUM for cell adhesion Zelladhäsion (DE-588)4314136-5 gnd Proteintyrosinphosphatase (DE-588)4311372-2 gnd |
| subject_GND | (DE-588)4314136-5 (DE-588)4311372-2 (DE-588)4113937-9 |
| title | Role of the novel protein tyrosine phosphatase AUM for cell adhesion |
| title_alt | Die Rolle des neuen Proteins "Tyrosin phosphatase" AUM für Zell-Adhäsion |
| title_auth | Role of the novel protein tyrosine phosphatase AUM for cell adhesion |
| title_exact_search | Role of the novel protein tyrosine phosphatase AUM for cell adhesion |
| title_full | Role of the novel protein tyrosine phosphatase AUM for cell adhesion von Ambrish Saxena |
| title_fullStr | Role of the novel protein tyrosine phosphatase AUM for cell adhesion von Ambrish Saxena |
| title_full_unstemmed | Role of the novel protein tyrosine phosphatase AUM for cell adhesion von Ambrish Saxena |
| title_short | Role of the novel protein tyrosine phosphatase AUM for cell adhesion |
| title_sort | role of the novel protein tyrosine phosphatase aum for cell adhesion |
| topic | Zelladhäsion (DE-588)4314136-5 gnd Proteintyrosinphosphatase (DE-588)4311372-2 gnd |
| topic_facet | Zelladhäsion Proteintyrosinphosphatase Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-65503 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026883678&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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