Interactions of smads and WW domains of proteins involved in TGF-beta signalling:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2011
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| Schlagworte: | |
| Online-Zugang: | Volltext Inhaltsverzeichnis |
| Beschreibung: | Nebentitel: Wechselwirkungen zwischen Smads und WW Domänen verschiedener TGF-ß Proteine |
| Beschreibung: | XXII, 141 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Interactions of smads and WW domains of proteins involved in TGF-beta signalling |c vorgelegt von Nina Görner |
| 246 | 1 | 3 | |a Wechselwirkungen zwischen Smads und WW-Domänen verschiedener TGF-Beta-Proteine |
| 264 | 1 | |c 2011 | |
| 300 | |a XXII, 141 S. |b Ill., graph. Darst. | ||
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| 500 | |a Nebentitel: Wechselwirkungen zwischen Smads und WW Domänen verschiedener TGF-ß Proteine | ||
| 502 | |a Berlin, Freie Univ., Diss., 2011 | ||
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Datensatz im Suchindex
| _version_ | 1804148948898676736 |
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| adam_text | IMAGE 1
TABLE O F CONTENTS
1 INTRODUCTION 1
1.1 TGF-FI SIGNALLING A N D SMADS 1
1.1.1 R-SMAD LINKER REGION 3
1.2 R-SMAD BINDING PROTEINS WITH W W DOMAINS 4
1.2.1 H E C T T Y P E E3-UBIQUITIN LIGASES 5
1.2.2 E3-UBIQUITIN LIGASE NEDD4L 7
1.2.3 PEPTIDYL-PROLYL ISOMERASE P I N L 8
1.3 I-SMAD7 BINDING PROTEINS WITH W W DOMAINS 9
1.3.1 TRANSCRIPTION CO-ACTIVATORS YAP A N D TAZ 10
2 AIMS O F THIS THESIS 1 1
3 METHODS AND MATERIALS 15
3.1 CLONING O F W W DOMAINS 17
3.1.1 CLONING OF T H E NEDD4L WW DOMAINS 1 7
3.1.2 CLONING OF T H E NEDD4L WW2 M U T A N T S 1 8
3.1.3 CLONING OF T H E NEDD4L WW2-WW3, WW1-WW2 A N D WW3-WW4 CONSTRUCTS
1 9
3.1.4 CLONING OF T H E NEDD4L WW2-WW3 P O I N T M U T A N T S 2 0
3.1.5 CLONING OF T H E P I N L W W DOMAIN 2 0
3.1.6 CLONING OF T H E YAP/TAZ W W DOMAINS 2 1
3.2 EXPRESSION AND PURIFICATION O F PROTEIN DOMAINS 2 1
3.2.1 TRANSFORMATION 2 1
3.2.2 RECOMBINANT PROTEIN EXPRESSION 2 2
3.2.3 CELL LYSIS 2 2
3.2.4 PROTEIN PURIFICATION 2 2
3.2.5 DETERMINATION OF PROTEIN CONCENTRATIONS 2 3
3.2.6 BUFFERS A N D SOLUTIONS/MATERIAL 1 2 4
3.3 PEPTIDE SYNTHESIS 2 5
3.3.1 MANUAL FMOC-SOLID P H A S E P E P T I D E SYNTHESIS 2 7
3.3.2 AUTOMATIC FMOC-SOLID P H A S E P E P T I D E SYNTHESIS 2 8
HTTP://D-NB.INFO/1026263085
IMAGE 2
TABLE O F C O N T E N T
3.3.3 SYNTHESIS OF PHOSPHOPEPTIDES 2 9
3.3.4 PEPTIDE PURIFICATION 2 9
3.3.5 MASS SPECTROMETRY 3 0
3.3.6 DETERMINATION OF PEPTIDE CONCENTRATIONS 3 1
3.3.7 BUFFERS A N D SOLUTIONS / MATERIAL II 3 1
3.4 ISOTHERMAL TITRATION CALORIMETRY 3 2
3.5 CHEMICAL LIGATION O F THE NEDD4L WW2-WW3 F R A G M E N T 3 4
3.5.1 CLONING OF T H E NEDD4L WW2-LINKER-CYS A N D CYS(GLY)-WW3
CONSTRUCTS 3 5
3.5.2 ACTIVATION OF T H E NEDD4L CYS(GLY)-WW3 DOMAIN 3 6
4 NMR SPECTROSCOPY 3 7
4.1 PRINCIPLES 3 9
4.2 ONE-DIMENSIONAL (ID) NMR EXPERIMENTS 41
4.3 TWO-DIMENSIONAL (2D) NMR EXPERIMENTS 4 2
4.3.1 NUCLEAR OVERHAUSER EFFECT 4 2
4.3.2 TOTAL CORRELATION SPECTROSCOPY 4 3
4.3.3 HETERONUCLEAR SINGLE Q U A N T U M COHERENCE 4 3
4.4 THE M A G N E T AND SIGNAL DETECTION 4 4
4.5 SAMPLE PREPARATION 4 4
4.6 PROTEIN STRUCTURE DETERMINATION 4 4
4.6.1 SEQUENTIAL RESONANCE ASSIGNMENT 4 5
4.6.2 STRUCTURE CALCULATION A N D REFINEMENT 4 6
5 RESULTS 5 1
5.1 INTERACTION O F THE R-SMAD3 LINKER AND W W DOMAINS 51
5.1.1 BINDING OF T H E S M A D 2 / 3 LINKER A N D NEDD4L WW DOMAINS 5 1
5.1.2 STRUCTURAL DETERMINATION OF T H E NEDD4L WW2 - SMAD3 PT[PY]
COMPLEX 5 6
5.1.3 THE SMAD3 PT179 P H O S P H A T E GROUP IS DIRECTLY INVOLVED IN
NEDD4L WW2 BINDING 5 8
5.1.4 THE PHOSPHORYLATED SMAD3 LINKER INTERACTS WITH T H E P I N L W W
DOMAIN 6 1
5.1.5 SMAD3 PT179P IS T H E PREFERRED PIN 1 W W BINDING SITE 6 2
5.1.6 THE SMAD3 PT179 P H O S P H A T E GROUP IS DIRECTLY INVOLVED IN P
I N L W W BINDING 6 4
5.1.7 STRUCTURAL DETERMINATION OF T H E P I N L WW - SMAD3 PT179 COMPLEX
6 6
5.1.8 P I N L W W BINDS T H E PS213P MOTIF OF T H E SMAD3 PS208-PS213
PEPTIDE 6 8
5.1.9 BINDING AFFINITY B E T W E E N SMAD3 A N D NEDD4L W W 2 / P I N L
WW D E P E N D S ON T H E PH 7 1
5.2 THE NEDD4L WW2-WW3 P A I R BINDS THE EXTEND SMAD3 LINKER 73
XX
IMAGE 3
TABLE OF CONTENTS
5.2.1 BINDING OF T H E NEDD4L WW1-WW2 P A I R A N D SMAD3 7 4
5.2.2 BINDING OF T H E NEDD4L WW3-WW4 P A I R A N D SMAD3 7 5
5.2.3 BINDING OF T H E NEDD4L WW2-WW3 P A I R A N D SMAD3 7 6
5.2.4 CHEMICAL LIGATION O F NEDD4L WW2 ( 2H15N) A N D NEDD4L W W 3 M
(15N) 8 0
5.2.5 STRUCTURE DETERMINATION OF T H E NEDD4L WW3 - SMAD3 PS-PS COMPLEX
8 4
5.3 BINDING O F THE I-SMAD7 LINKER AND W W DOMAINS 8 7
5.3.1 SMAD7 LINKER BINDS NEDD4L WW2 DOMAIN WITH HIGH AFFINITY 8 7
5.3.2 STRUCTURAL DETERMINATION OF T H E NEDD4L WW2 - SMAD7[PY] COMPLEX 9
0
5.3.3 SMAD7 LINKER INTERACTS WITH YAP/TAZ W W DOMAINS 9 5
5.3.4 STRUCTURAL DETERMINATION OF T H E TAZ WW - SMAD7[PY] COMPLEX 9 7
6 DISCUSSION 1 0 3
6.1 INTERACTION O F S M A D 3 AND THE W W DOMAINS OFNEDD4L AND PINL 1 0
3
6.1.1 SELECTIVITY IN T H E NEDD4L - SMAD3 INTERACTION 103
6.1.2 ROLE OF P I N L IN SMAD3 RECOGNITION 1 0 5
6.1.3 31P NMR EXPERIMENTS OF PHOSPHORYLATED SMAD3 PEPTIDES 1 0 6
6.1.4 SMAD3 - W W DOMAIN BINDING IS AFFECTED BY PH CHANGES 1 0 6
6.1.5 COOPERATIVITY O F W W DOMAINS IN NEDD4L - SMAD3 INTERACTION 1 0 7
6.2 INTERACTION O F S M A D 7 AND THE W W DOMAINS OFNEDD4L AND TAZ 1 1 0
6.2.1 BINDING OF SMAD7 A N D NEDD4L 1 1 0
6.2.2 BINDING OF SMAD7 A N D TAZ I L L
7 APPENDIX 1 1 5
7.1 AMINO ACIDS 1 1 5
7.2 NEDD4L 116
7.2.1 FULL-LENGTH HNEDD4L PROTEIN SEQUENCE 1 1 6
7.2.2 PATTERN OF SECONDARY S T R U C T U R E NOES (NEDD4L W W DOMAINS) 1
1 6
7.2.3 NMR TITRATIONS: NEDD4L W W DOMAINS + S M A D 2 / 3 PT[PY] 1 1 7
7.3 PINL 1 1 8
7.3.1 FULL-LENGTH H P I N L PROTEIN SEQUENCE 1 1 8
7.3.2 PATTERN OF SECONDARY S T R U C T U R E NOES ( P I N L W W DOMAIN)
119
7.4 YAPANDTAZ. 1 1 9
7.4.1 FULL-LENGTH HYAP PROTEIN SEQUENCE 1 1 9
7.4.2 FULL-LENGTH HTAZ PROTEIN SEQUENCE 119
7.4.3 PATTERN OF SECONDARY S T R U C T U R E NOES (TAZ W W DOMAIN) 1 2 0
XXI
IMAGE 4
TABLE OF CONTENTS
7.5 STRUCTURE STATISTICS 1 2 0
7.6 PROTON NOMENCLATURE F O R NMR ASSIGNMENT. 121
GLOSSARY 1 2 3
LIST OF FIGURES 127
LIST O F TABLES 133
REFERENCES 135
XXII
|
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| author_facet | Görner, Nina |
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| ctrlnum | (OCoLC)918445268 (DE-599)BVBBV039970491 |
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| dewey-ones | 570 - Biology |
| dewey-raw | 570 |
| dewey-search | 570 |
| dewey-sort | 3570 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| physical | XXII, 141 S. Ill., graph. Darst. |
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| spelling | Görner, Nina Verfasser aut Interactions of smads and WW domains of proteins involved in TGF-beta signalling vorgelegt von Nina Görner Wechselwirkungen zwischen Smads und WW-Domänen verschiedener TGF-Beta-Proteine 2011 XXII, 141 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Nebentitel: Wechselwirkungen zwischen Smads und WW Domänen verschiedener TGF-ß Proteine Berlin, Freie Univ., Diss., 2011 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:kobv:188-fudissthesis000000035451-8 http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000035451 kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024828049&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Görner, Nina Interactions of smads and WW domains of proteins involved in TGF-beta signalling |
| subject_GND | (DE-588)4113937-9 |
| title | Interactions of smads and WW domains of proteins involved in TGF-beta signalling |
| title_alt | Wechselwirkungen zwischen Smads und WW-Domänen verschiedener TGF-Beta-Proteine |
| title_auth | Interactions of smads and WW domains of proteins involved in TGF-beta signalling |
| title_exact_search | Interactions of smads and WW domains of proteins involved in TGF-beta signalling |
| title_full | Interactions of smads and WW domains of proteins involved in TGF-beta signalling vorgelegt von Nina Görner |
| title_fullStr | Interactions of smads and WW domains of proteins involved in TGF-beta signalling vorgelegt von Nina Görner |
| title_full_unstemmed | Interactions of smads and WW domains of proteins involved in TGF-beta signalling vorgelegt von Nina Görner |
| title_short | Interactions of smads and WW domains of proteins involved in TGF-beta signalling |
| title_sort | interactions of smads and ww domains of proteins involved in tgf beta signalling |
| topic_facet | Hochschulschrift |
| url | http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000035451 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024828049&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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