Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells:
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| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2011
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-146738 Inhaltsverzeichnis |
| Beschreibung: | II, 164 S. Ill., graph. Darst. 1 CD-ROM (12 cm) |
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MARC
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| 245 | 1 | 0 | |a Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells |c Melanie Schulz |
| 264 | 1 | |c 2011 | |
| 300 | |a II, 164 S. |b Ill., graph. Darst. |e 1 CD-ROM (12 cm) | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Univ., Diss., 2011 | ||
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| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-025212892 | ||
Datensatz im Suchindex
| _version_ | 1804149399447666688 |
|---|---|
| adam_text | IMAGE 1
CONTENTS
ABSTRACT I
1 INTRODUCTION 1
1.1 2,3,7,8-TETRACHLORODIBENZO-J5-DIOXIN 1
1.1.1 THE GENOMIC ACTIONS OF TCDD 1
1.1.2 THE NONGENOMIC ACTIONS OF TCDD 2
1.2 PHOSPHOPROTEOMICS 4
1.2.1 PHOSPHOPEPTIDE ENRICHMENT STRATEGIES 5
1.2.2 SHOTGUN PROTEOMICS 8
1.2.3 QUANTITATIVE PHOSPHOPROTEOMICS 11
1.3 THE AIM OF THE PRESENT STUDY 12
2 MATERIAL A N D M E T H O D S 15
2.1 MATERIALS . . . 15
2.1.1 SOFTWARE 16
2.2 METHODS 16
2.2.1 CELL CULTURE 16
2.2.2 SAMPLE PREPARATION 18
2.2.3 MASS SPECTROMETRIC METHODS 21
2.2.4 BIOINFORMATICS AND BIOSTATISTICS ANALYSIS 22
2.2.5 WESTERN BLOT ANALYSIS 24
2.2.6 ETHOXYRESORUFIN O-DEETHYLASE (EROD) ASSAY 25
3 RESULTS 2 7
3.1 SILAC LABELLING 28
3.2 SIM AC FOR PHOSPHOPEPTIDE ENRICHMENT 32
3.2.1 APPLYING SIMAC TO STANDARD PEPTIDES 32
HTTP://D-NB.INFO/1025274962
IMAGE 2
3.2.2 APPLYING SIM AC TO COMPLEX SAMPLES . . 36
3.3 HILIC AS A PREFRACTIONATION METHOD 37
3.4 THE PHOSPHOPROTEOME OF 5L CELLS 40
3.4.1 IDENTIFIED PHOSPHORYLATION SITES 40
3.5 MULTIPLE TESTING FOR FINDING STATISTICALLY SIGNIFICANTLY REGULATED
PHOSPHO- . PEPTIDES 43
3.5.1 NORMALISATION 43
3.5.2 CORRELATION 45
3.5.3 STATISTICAL ANALYSIS 45
3.6 PEPTIDES WITH ALTERED PHOSPHORYLATION AFTER TREATMENT OF THE 5L
CELLS WITH
TCDD 49
3.6.1 TIME COURSE OF THE INDUCTION OF CYP1A1 49
3.6.2 PEPTIDES WITH ALTERED PHOSPHORYLATION AFTER 30MIN TCDD TREATMENT
51 3.6.3 PEPTIDES WITH ALTERED PHOSPHORYLATION AFTER 1 HOUR TCDD
TREATMENT 53 3.6.4 PEPTIDES WITH ALTERED PHOSPHORYLATION AFTER 2 HOURS
TCDD TREATMENT 58 3.6.5 SUMMARY OF THE IDENTIFIED PEPTIDES WITH ALTERED
PHOSPHORYLATION AF
TER TCDD TREATMENT 61
3.7 MECHANISM OF THE CALCIUM DEPENDENCE OF TCDD-INDUCED GENE ACTIVATION
IN 5L CELLS 67
4 DISCUSSION 71
4.1 METHODOLOGICAL ASPECTS 71
4.1.1 SILAC LABELLING 71
4.1.2 A COMPREHENSIVE PHOSPHOPROTEOMIC ANALYSIS? 73
4.1.3 ENRICHMENT OF PHOSPHOPEPTIDES 74
4.1.4 STATISTICS 75
4.1.5 REPRODUCIBILITY 76
4.2 THE IDENTIFIED TCDD-INDUCED ALTERATIONS IN PROTEIN PHOSPHORYLATION .
. . . 78 4.2.1 PROTEINS PREVIOUSLY ASSOCIATED WITH NONGENOMIC ACTIONS OF
TCDD . 79 4.2.2 AHR ( AHR ) AND ARNT ( ARNT ) 80
4.2.3 REGULATORS OF THE TRANSCRIPTIONAL MACHINERY INVOLVED IN
AHR/ARNTMEDIATED GENE ACTIVATION 81
4.2.4 REGULATORS OF SMALL GTPASES OF THE RAS SUPERFAMILY 86
4.2.5 UBX DOMAIN-CONTAINING PROTEINS 91
4.2.6 MISCELLANEOUS PROTEINS 93
IMAGE 3
5 CONCLUSIONS
A P P E N D I X A TABLES
A P P E N D I X B FIGURES
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| dewey-ones | 571 - Physiology & related subjects |
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| discipline | Biologie |
| format | Thesis Book |
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| spelling | Schulz, Melanie 1980- Verfasser (DE-588)1024983757 aut Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells Melanie Schulz 2011 II, 164 S. Ill., graph. Darst. 1 CD-ROM (12 cm) txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2011 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-146738 http://edoc.ub.uni-muenchen.de/14673/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-146738 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025212892&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schulz, Melanie 1980- Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells |
| subject_GND | (DE-588)4113937-9 |
| title | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells |
| title_auth | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells |
| title_exact_search | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells |
| title_full | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells Melanie Schulz |
| title_fullStr | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells Melanie Schulz |
| title_full_unstemmed | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells Melanie Schulz |
| title_short | Quantitative proteomic analysis of early phosphorylation changes induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in 5L rat hepatoma cells |
| title_sort | quantitative proteomic analysis of early phosphorylation changes induced by 2 3 7 8 tetrachlorodibenzo p dioxin in 5l rat hepatoma cells |
| topic_facet | Hochschulschrift |
| url | http://edoc.ub.uni-muenchen.de/14673/ https://nbn-resolving.org/urn:nbn:de:bvb:19-146738 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025212892&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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