Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120709-1096625-1-4 Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. Sprache |
| Beschreibung: | VIII, 119 Bl. graph. Darst. |
Internformat
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| 100 | 1 | |a Hirschberg, Verena |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines |c Verena Hirschberg |
| 264 | 1 | |c 2012 | |
| 300 | |a VIII, 119 Bl. |b graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Zsfassung in dt. Sprache | ||
| 502 | |a München, Techn. Univ., Diss., 2012 | ||
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| 650 | 0 | 7 | |a Enzymaktivität |0 (DE-588)4152470-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Zelllinie |0 (DE-588)4231928-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Protein-Protein-Wechselwirkung |0 (DE-588)4745193-2 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Mitochondrial Uncoupling Protein |0 (DE-588)4599869-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Wärmeerzeugung |0 (DE-588)4188852-2 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 689 | 0 | 1 | |a Zellkultur |0 (DE-588)4067547-6 |D s |
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| 856 | 4 | |u https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120709-1096625-1-4 |x Resolving-System | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025193790&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
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Datensatz im Suchindex
| _version_ | 1804149372092416000 |
|---|---|
| adam_text | IMAGE 1
I
ABBREVIATIONS IV
FIGURES VI
TABLES VIII
1 INTRODUCTION 1
1.1 UNCOUPLED RESPIRATION AND ITS PHYSIOLOGICAL SIGNIFICANCE 1
1.2 MOLECULAR ASPECTS O F UCP1 FUNCTION 3
1.3 BENEFICIAL EFFECTS OF INCREASED UNCOUPLING AND HUMAN BAT 5
1.4 TEST SYSTEMS FOR UCP1 FUNCTION 6
1.5 CIDEA 8
2 MATERIAL AND METHODS 10
2.1 MOLECULAR BIOLOGY 10
2.1.1 RNA ISOLATION, CDNA SYNTHESIS, QRT PCR 10
2.1.2 PROTEIN EXTRACTION AND QUANTITFICATION 1 1
2.1.3 IMMUNOLOGICAL DETECTION O F PROTEINS 13
2.2 LIPOSOME ANALYSIS 16
2.2.1 PURIFICATION AND RECONSTITUTION O F MOUSE UCP1 17
2.2.2 HETEROLOGOUS EXPRESSION AND PURIFICATION O F CIDEA IN E.COLI 18
2.2.3 ANALYSIS O F UCP1 FUNCTION IN LIPOSOMES 20
2.3 CELL CULTURE 2 1
2.3.1 CULTIVATION OF HEK293 AND HEK293 UCP1 CELLS 2 1
2.3.2 TRANSIENT OR STABLE CALCIUM-PHOSPHATE-MEDIATED TRANSFECTION 22
2.3.3 CULTIVATION OF IMMORTALISED BROWN PREADIPOCYTES 23
2.3.4 ANALYSIS O F LIPID STORAGE AND LIPOLYSIS 26
2.4 ISOLATION OF MITOCHONDRIA 28
2.4.1 ISOLATION O F HEK293 MITOCHONDRIA 28
2.4.2 ISOLATION O F MITOCHONDRIA FROM CULTURED BROWN ADIPOCYTES 29
2.4.3 ISOLATION O F MITOCHONDRIA FROM BROWN ADIPOSE TISSUE 29
2.5 MEASUREMENTS OF MITOCHONDRIAL AND CELLULAR BIOENERGETICS 30
2.5.1 DETERMINATION O F PROTON LEAK KINETICS O F ISOLATED MITOCHONDRIA
30
2.5.2 ANALYSIS O F OXYGEN CONSUMPTION O F TRYPSINISED CELLS 37
2.5.3 ANALYSIS O F OXYGEN CONSUMPTION O F ADHERENTLY GROWING CELLS 37
2.6 STATISTICAL ANALYSIS 39
HTTP://D-NB.INFO/1032095032
IMAGE 2
II
3 RESULTS 41
3.1 UCP1 ACTIVITY IN LIPOSOMES, HEK293 CELLS, IMMORTALISED BROWN
ADIPOCYTES AND ISOLATED
BAT MITOCHONDRIA 4 1
3.1.1 UCP1 ACTIVITY IN LIPOSOMES 4 1
3.1.2 UCP1 ACTIVITY IN HEK293 CELLS 43
3.1.3 UCP1 ACTIVITY IN IMMORTALISED BROWN ADIPOCYTES (BFC) 54
3.1.4 UCP1 ACTIVITY IN MITOCHONDRIA ISOLATED FROM MOUSE BAT 6 1
3.1.5 COMPARISON O F UCPL-DEPENDENT PROTON LEAK AND CATALYTIC ACTIVITY
IN DIFFERENT TEST SYSTEMS 63
3.2 INTERACTION OF CIDEA AND UCP1 IN DIFFERENT TEST SYSTEMS 67
3.2.1 GENERATION O F AN ANTIBODY FOR CIDEA DETECTION 67
3.2.2 CIDEA EXPRESSION IN MOUSE TISSUES 69
3.2.3 INTERACTION O F CIDEA AND UCP1 RECONSTITUTED INTO LIPOSOMES 69
3.2.4 INTERACTION O F CIDEA AND UCP1 IN HEK293 CELLS 72
3.2.5 OVEREXPRESSION O F CIDEA IN IMMORTALISED BROWN ADIPOCYTES 75
4 DISCUSSION 80
4.1 SUITABILITY OF HEK293 CELLS AND BFC AS TEST SYSTEMS FOR UCP FUNCTION
80
4.1.1 EXPRESSION O F UCP1 IN HEK293 UCP1 AND BFC 80
4.1.2 REGULATION O F UCP1 ACTIVITY IN HEK293 UCP1 AND BFC MITOCHONDRIA
84
4.1.3 UCP1 IN HEK293 UCP1 AND BFC: NO UNCOUPLING ARTEFACT OR
CONTRIBUTION T O BASAL LEAK 85
4.1.4 BASAL ACTIVITY O F UCP1 IN THE CELLULAR CONTEXT 86
4.1.5 SECONDARY EFFECTS OF UCP1 EXPRESSION IN HEK293 AND BFC 88
4.1.6 CATALYTIC ACTIVITY OF UCP1 IN THE DIFFERENT TEST SYSTEMS 90
4.1.7 POTENTIAL O F THE HEK293 CELLS AND BFC FOR UCP RESEARCH 92
4.2 INTERACTION OF UCP1 AND CIDEA IN DIFFERENT TEST SYSTEMS 94
4.2.1 CIDEA, A MITOCHONDRIAL PROTEIN? 94
4.2.2 NO EFFECT O F CIDEA EXPRESSION ON UCP1 ACTIVITY IN THE TEST
SYSTEMS 94
4.2.3 CIDEA AS A REGULATOR O F LIPID METABOLISM? 96
4.2.4 INTERACTION O F CIDEA AND UCP1 VIA FATTY ACID LEVELS 97
5 SUMMARY AND CONCLUSION 99
6 REFERENCES 101
7 APPENDIX 110
7.1 MATERIAL 110
7.2 QRT PCR 112
7.3 VECTORS 113
7.4 SEAHORSE PROTOCOL 113
IMAGE 3
ILL
8 KURZFASSUNG (ABSTRACT) 114
9 ERKLARUNG 115
10 CURRICULUM VITAE 116
11 DANKSAGUNG 119
|
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| ctrlnum | (OCoLC)812198097 (DE-599)BVBBV040339501 |
| discipline | Biologie Chemie |
| format | Thesis Book |
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| spelling | Hirschberg, Verena Verfasser aut Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines Verena Hirschberg 2012 VIII, 119 Bl. graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache München, Techn. Univ., Diss., 2012 Zellkultur (DE-588)4067547-6 gnd rswk-swf Enzymaktivität (DE-588)4152470-6 gnd rswk-swf Zelllinie (DE-588)4231928-6 gnd rswk-swf Protein-Protein-Wechselwirkung (DE-588)4745193-2 gnd rswk-swf Mitochondrial Uncoupling Protein (DE-588)4599869-3 gnd rswk-swf Wärmeerzeugung (DE-588)4188852-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Zelllinie (DE-588)4231928-6 s Zellkultur (DE-588)4067547-6 s Mitochondrial Uncoupling Protein (DE-588)4599869-3 s Wärmeerzeugung (DE-588)4188852-2 s Enzymaktivität (DE-588)4152470-6 s Protein-Protein-Wechselwirkung (DE-588)4745193-2 s b DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20120709-1096625-1-4 http://mediatum.ub.tum.de/node?id=1096625 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120709-1096625-1-4 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025193790&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hirschberg, Verena Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines Zellkultur (DE-588)4067547-6 gnd Enzymaktivität (DE-588)4152470-6 gnd Zelllinie (DE-588)4231928-6 gnd Protein-Protein-Wechselwirkung (DE-588)4745193-2 gnd Mitochondrial Uncoupling Protein (DE-588)4599869-3 gnd Wärmeerzeugung (DE-588)4188852-2 gnd |
| subject_GND | (DE-588)4067547-6 (DE-588)4152470-6 (DE-588)4231928-6 (DE-588)4745193-2 (DE-588)4599869-3 (DE-588)4188852-2 (DE-588)4113937-9 |
| title | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines |
| title_auth | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines |
| title_exact_search | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines |
| title_full | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines Verena Hirschberg |
| title_fullStr | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines Verena Hirschberg |
| title_full_unstemmed | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines Verena Hirschberg |
| title_short | Analysis of uncoupling protein 1 and CideA function in two mammalian cell lines |
| title_sort | analysis of uncoupling protein 1 and cidea function in two mammalian cell lines |
| topic | Zellkultur (DE-588)4067547-6 gnd Enzymaktivität (DE-588)4152470-6 gnd Zelllinie (DE-588)4231928-6 gnd Protein-Protein-Wechselwirkung (DE-588)4745193-2 gnd Mitochondrial Uncoupling Protein (DE-588)4599869-3 gnd Wärmeerzeugung (DE-588)4188852-2 gnd |
| topic_facet | Zellkultur Enzymaktivität Zelllinie Protein-Protein-Wechselwirkung Mitochondrial Uncoupling Protein Wärmeerzeugung Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1096625 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120709-1096625-1-4 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025193790&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT hirschbergverena analysisofuncouplingprotein1andcideafunctionintwomammaliancelllines |