Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry: Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2010
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| Schlagworte: | |
| Online-Zugang: | kostenfrei https://nbn-resolving.org/urn:nbn:de:bvb:19-155369 Inhaltsverzeichnis |
| Beschreibung: | 105 Bl. Ill., graph. Darst. |
Internformat
MARC
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| 035 | |a (DE-599)BVBBV040893257 | ||
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| 100 | 1 | |a Dulla, Kalyana Chakravarthi |d 1982- |e Verfasser |0 (DE-588)103253141X |4 aut | |
| 245 | 1 | 0 | |a Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry |b Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages |c von Kalyana Chakravarthi Dulla |
| 264 | 1 | |c 2010 | |
| 300 | |a 105 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Univ., Diss., 2010 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:19-155369 |
| 856 | 4 | 1 | |u http://edoc.ub.uni-muenchen.de/15536/ |x Verlag |z kostenfrei |3 Volltext |
| 856 | 4 | |u https://nbn-resolving.org/urn:nbn:de:bvb:19-155369 |x Resolving-System | |
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| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-025872860 | ||
Datensatz im Suchindex
| _version_ | 1804150176492814336 |
|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS
SUMMARY 7
1.0 INTRODUCTION 8
1.1 CELL CYCLE 8
1.2 MITOSIS 8
1.2.1 SPINDLE ASSEMBLY CHECKPOINT 10
1.2.2 THE MOLECULAR MECHANISM O F THE SAC 10
1.3 REGULATION O F KINASES BY PHOSPHORYLATION 12
1.4 MASS SPECTROMETRY IN BIOLOGY 13
1.5 MASS SPECTROMETRIC INSTRUMENTATION 14
1.5.1 ION SOURCE 14
1.5.2 MASS ANALYZERS 15
1.5.2.1 LINEAR QUADRUPOLE ION TRAP 16
1.5.2.2 THE ELECTROSTATIC TRAP OR ORBITRAP 19
1.5.3 DETECTORS 20
1.6 METHODS FOR QUANTITATION 20
1.7 PHOSPHOPROTEOMICS 22
1.8 BIOINFORMATIC TOOLS FOR THE ANALYSIS OF MASS SPECTROMETRY DATA 23
1.8.1 GENE ONTOLOGY 23
1.8.2 PATHWAY ANALYSIS 24
1.8.3 INTERACTION NETWORKS 25
2.0 RESULTS (PART 1): PHOSPHORYLATION DYNAMICS OF PROTEIN KINASES ACROSS
MITOSIS 26
2.1 INTRODUCTION 26
2.2 EXPERIMENTAL STRATEGY 26
4
HTTP://D-NB.INFO/1033288179
IMAGE 2
2.3 LARGE SCALE SYNCHRONIZATION O F HELA CELLS IN DIFFERENT MITOTIC
STAGES 27
2.4 IDENTIFICATION OF PROTEINS AND PHOSPHORYLATION SITES 32
2.5 MITOTIC DYNAMICS OF KINASE PHOSPHORYLATION SITES 34
2.6 NETWORK ANALYSIS 41
2.7 MAPK PATHWAYS ARE ACTIVATED DURING LATE MITOSIS 43
2.8 CHANGES MADE DURING MITOTIC ENTRY ARE RESTORED AT THE END OF MITOSIS
45
2.9 REGULATION OF NON-KINASES IN MITOSIS 46
3.0 RESULTS (PART 2): CHARACTERIZATION OF FUNCTION OF BUBRL AND BUBL
KINASE PHOSPHORYLATION..50
3.1 INTRODUCTION 50
3.2 IDENTIFICATION OF BUBRL PHOSPHORYLATION SITES IN MITOSIS 51
3.3 CDKL PHOSPHORYLATES S543, S670, AND SI043 IN-VITRO 54
3.4 BUBRL-5A MUTANT EXPRESSING CELLS EXHIBIT SEVERE CONGRESSION DEFECTS
55
3.5 BUBL PHOSPHORYLATION IN MITOSIS 58
3.6 INCREASING THE SEQUENCE COVERAGE OF BUBL BY USING MULTIPLE PROTEASES
59
3.7 IDENTIFICATION OF AUTO-PHOSPHORYLATION SITES ON BUBL 60
3.8 IDENTIFICATION OF PHOSPHORYLATION ON BUBL INTERACTORS 64
4.0 DISCUSSION 65
4.1 SMALL-MOLECULE KINASE INHIBITOR BASED ENRICHMENT OF THE KINOME 65
4.2 HELA CELL SYNCHRONIZATION 66
4.3 IDENTIFICATION OF PROTEIN KINASES 67
4.4 GLOBAL VIEW OF MITOTIC KINOME 68
4.5 REGULATION OF BUBRL FUNCTION BY PHOSPHORYLATION 69
4.6 REGULATION OF SAC AND CHROMOSOME CONGRESSION BY DIFFERENT MOTIFS OF
BUBRL 70
4.7 AUTOPHOSPHORYLATION AND SUBSTRATES OF BUBL KINASE 70
4.8 ADVANCES IN MS INSTRUMENTATION AND DATA ANALYSIS TOOLS MAKE LARGE
SCALE PROTEOMIC
STUDIES POSSIBLE 72
5
IMAGE 3
5.0 EXPERIMENTAL PROCEDURES: 74
5.1 ANTIBODIES 74
5.2 CELL CULTURE, S1LAC LABELING AND SYNCHRONIZATION 74
5.3 IMMUNOFLUORESCENCE STAINING O F CELLS 75
5.4 KINASE ENRICHMENT 75
5.5 IN-GEL PROTEIN DIGESTION 75
5.6 IN-SOLUTION PROTEIN DIGESTION 76
5.7 STRONG-CATION EXCHANGE (SCX) 77
5.8 PHOSPHOPEPTIDE ENRICHMENT AND DESALTING 77
5.9 NANO-LC-MS/MS ANALYSIS 77
5.10 DATA PROCESSING AND ANALYSIS 78
5.11 INTERACTION NETWORKS AND FUNCTIONAL CLUSTERING 79
5.12 CELL LYSIS AND CO-IMMUNOPRECIPITATION 79
5.13 KINASE ASSAY 80
5.14 PEPTIDE ARRAY SYNTHESIS AND SPOTS BLOTTING 81
6.0 APPENDIX 82
6.1 PPI NETWORK FOR PROTEINS WITH UPREGULATED PHOSPHORYLATION SITES 82
6.2 PPI NETWORK FOR PROTEINS WITH DOWNREGULATED PHOSPHORYLATION SITES 83
6.3 SUPPORTING DATA AVAILABLE AT TRANCHE 84
ABBREVIATIONS 85
REFERENCES 87
ACKNOWLEDGEMENTS 102
PUBLICATIONS 103
CURRICULUM VITAE 104
6
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| indexdate | 2024-07-10T00:34:46Z |
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| language | English |
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| spelling | Dulla, Kalyana Chakravarthi 1982- Verfasser (DE-588)103253141X aut Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages von Kalyana Chakravarthi Dulla 2010 105 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2010 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-155369 http://edoc.ub.uni-muenchen.de/15536/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-155369 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025872860&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Dulla, Kalyana Chakravarthi 1982- Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages |
| subject_GND | (DE-588)4113937-9 |
| title | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages |
| title_auth | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages |
| title_exact_search | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages |
| title_full | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages von Kalyana Chakravarthi Dulla |
| title_fullStr | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages von Kalyana Chakravarthi Dulla |
| title_full_unstemmed | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages von Kalyana Chakravarthi Dulla |
| title_short | Analysis of mitotic phosphoproteomes using SILAC based mass spectrometry |
| title_sort | analysis of mitotic phosphoproteomes using silac based mass spectrometry focus on the site specific phosphorylation dynamics of kinases through different mitotic stages |
| title_sub | Focus on the site-specific phosphorylation dynamics of kinases through different mitotic stages |
| topic_facet | Hochschulschrift |
| url | http://edoc.ub.uni-muenchen.de/15536/ https://nbn-resolving.org/urn:nbn:de:bvb:19-155369 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025872860&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT dullakalyanachakravarthi analysisofmitoticphosphoproteomesusingsilacbasedmassspectrometryfocusonthesitespecificphosphorylationdynamicsofkinasesthroughdifferentmitoticstages |