Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Heidelberg
2022
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | v, 170 Seiten Illustrationen, Diagramme 30 cm |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV048918064 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | t | ||
| 008 | 230428s2022 gw a||| m||| 00||| eng d | ||
| 015 | |a 23,H03 |2 dnb | ||
| 016 | 7 | |a 1278052933 |2 DE-101 | |
| 020 | |c Broschur | ||
| 035 | |a (OCoLC)1372146732 | ||
| 035 | |a (DE-599)DNB1278052933 | ||
| 040 | |a DE-604 |b ger |e rda | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c XA-DE | ||
| 049 | |a DE-355 | ||
| 084 | |8 1\p |a 570 |2 23sdnb | ||
| 084 | |a 570 |2 sdnb | ||
| 100 | 1 | |a Bosch, Katharina |d 1989- |e Verfasser |0 (DE-588)1278718362 |4 aut | |
| 245 | 1 | 0 | |a Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |c presented by M.Sc. Katharina Bosch |
| 264 | 1 | |a Heidelberg |c 2022 | |
| 300 | |a v, 170 Seiten |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Ruperto Carola University Heidelberg, Germany |d 2022 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1278052933/04 |3 Inhaltsverzeichnis |
| 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=034182159&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-034182159 | ||
| 883 | 2 | |8 1\p |a dnb |d 20230215 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
Datensatz im Suchindex
| _version_ | 1804185095752384512 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
ACKNOWLEDGEMENTS
1
SUMMARY
3
ZUSAMMENFASSUNG
5
1.
INTRODUCTION
11
1.1.
SOURCES
AND
REGULATION
OF
OXIDANTS
........................................................
11
1.1.1.
HYDROGEN
PEROXIDE
AND
OTHER
BIOLOGICALLY
RELEVANT
OXIDANTS
.
.
12
1.1.2.
REGULATION
OF
OXIDANTS
................................................................
15
1.1.3.
METAL-BASED
OXIDANT
SCAVENGERS
.................................................
15
1.1.4.
THE
GLUTATHIONE
SYSTEM
.............................................................
15
1.1.5.
THE
THIOREDOXIN
SYSTEM
.............................................................
16
1.2.
PROTEIN
THIOL
OXIDATION
AND
SIGNALLING
...................................................
17
1.3.
ROGFP2
....................................................................................................
19
1.4.
YEAST
AS
A
MODEL
ORGANISM
FOR
REDOX
BIOLOGY
...........................................
21
1.5.
OBJECTIVE
.....................................................................................................
24
2.
MATERIALS
AND
METHODS
29
2.1.
MATERIALS
.....................................................................................................
29
2.1.1.
CHEMICALS
AND
BIOCHEMICALS
..........................................................
31
2.1.2.
KITS,
ENZYMES
AND
OTHERS
.............................................................
31
2.1.3.
ANTIBODIES
.....................................................................................
32
2.1.4.
OLIGONUCLEOTIDES
AND
PLASMIDS
.......................................................
33
2.1.5.
BUFFERS
AND
MEDIA
.........................................................................
42
2.1.6.
BIOLOGICAL
MATERIAL
.........................................................................
43
2.1.7.
SOFTWARE
...............................................................
44
2.2.
METHODS
........................................................................................................
45
2.2.1.
YEAST
GROWTH
CONDITIONS
................................................................
45
2.2.2.
TRANSFORMATION
OF
YEAST
................................................................
46
7
2.2.3.
LIBRARY
CONSTRUCTION
AND
VALIDATION
...............................................
47
2.2.4.
ADDITIONAL
YEAST
STRAINS
..................................................................
48
2.2.5.
LIBRARY
MEASUREMENT
........................................................................
48
2.2.6.
DATA
ANALYSIS
....................................................................................
49
2.2.7.
PLATE
READER
MEASUREMENTS
.......................................................
49
2.2.8.
MICROSCOPY
........................................................................................
50
2.2.9.
CELL
LYSIS,
SDS-PAGE,
AND
IMMUNOBLOTTING
...................
YY
.
.
.
50
2.2.10.
COVALENT
MODIFICATION
WITH
METHOXYPOLYETHYLENEGLYCOL
MALEIMIDE
(PEG-MAL)
....................................................................
50
2.2.11.
FLOW
CYTOMETRY
...............................................................................
51
2.2.12.
METABOLIC
SHIFT
EXPERIMENT
OF
SELECTED
STRAINS
...........................
51
2.2.13.
HYPOXIA
MEASUREMENTS
...................................................................
51
2.2.14.
ASNL-ROGFP2
MICROSCOPY
AND
PLATE
READER
ASSAY
......................
52
3.
RESULTS
53
3.1.
LIBRARY
CONSTRUCTION,
MEASUREMENT
AND
ANALYSIS
.........................................
53
3.2.
IDENTIFICATION
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
...........................
.
63
3.3.
IDENTIFICATION
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
USING
AN
ALTERNATIVE
APPROACH
.........................................................................................................
73
3.4.
FUSION
PROTEIN
SPECIFIC
REDOX
DIFFERENCES
ACROSS
DIFFERENT
METABOLIC
CONDITIONS
......................................................................................................
81
3.5.
OXIDATION
OF
ASNL-ROGFP2
CORRELATES
WITH
THE
FORMATION
OF
CYTOOPHIDIAE
...................................................................................................
86
4.
DISCUSSION
93
4.1.
CHALLENGES
OF
C-TERMINAL
TAGGING
IN
GENERAL
...........................................
93
4.2.
CHALLENGES
OF
ENDOGENOUS
C-TERMINAL
TAGGING
WITH
ROGFP2
...................
94
4.3.
GENERAL
OBSERVATIONS
...................................................................................
96
4.4.
LOCAL
AND
CONTEXT
SPECIFIC
ROGFP2
OXIDATION
...........................................
97
4.5.
POSSIBLE
OXIDATION
OF
ROGFP2
ASSOCIATED
PROTEINS
................................
100
4.6.
CONCLUSIONS
AND
FUTURE
DIRECTIONS
............................................................
101
5.
REFERENCES
107
8
APPENDICES
121
A.
APPENDIX
123
A.1.
LIST
OF
ROGFP2
FUSION
PROTEINS
FOUND
TO
BE
OXIDISED
USING
THE
R/R
APPROACH
..............................................................................................
123
A.
2.
GO
ENRICHMENT
INCLUDING
SEMANTIC
SIMILARITY
CLUSTERING
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
(R/R
APPROACH)
.......................................
126
A.
3.
LIST
OF
ROGFP2
FUSION
PROTEINS
FOUND
TO
BE
OXIDISED
USING
THE
485/485
APPROACH
..............................................................................................
131
A.
4.
GO
ENRICHMENT
INCLUDING
SEMANTIC
SIMILARITY
CLUSTERING
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
(485/485
APPROACH)
.................................
148
9
|
| adam_txt |
TABLE
OF
CONTENTS
ACKNOWLEDGEMENTS
1
SUMMARY
3
ZUSAMMENFASSUNG
5
1.
INTRODUCTION
11
1.1.
SOURCES
AND
REGULATION
OF
OXIDANTS
.
11
1.1.1.
HYDROGEN
PEROXIDE
AND
OTHER
BIOLOGICALLY
RELEVANT
OXIDANTS
.
.
12
1.1.2.
REGULATION
OF
OXIDANTS
.
15
1.1.3.
METAL-BASED
OXIDANT
SCAVENGERS
.
15
1.1.4.
THE
GLUTATHIONE
SYSTEM
.
15
1.1.5.
THE
THIOREDOXIN
SYSTEM
.
16
1.2.
PROTEIN
THIOL
OXIDATION
AND
SIGNALLING
.
17
1.3.
ROGFP2
.
19
1.4.
YEAST
AS
A
MODEL
ORGANISM
FOR
REDOX
BIOLOGY
.
21
1.5.
OBJECTIVE
.
24
2.
MATERIALS
AND
METHODS
29
2.1.
MATERIALS
.
29
2.1.1.
CHEMICALS
AND
BIOCHEMICALS
.
31
2.1.2.
KITS,
ENZYMES
AND
OTHERS
.
31
2.1.3.
ANTIBODIES
.
32
2.1.4.
OLIGONUCLEOTIDES
AND
PLASMIDS
.
33
2.1.5.
BUFFERS
AND
MEDIA
.
42
2.1.6.
BIOLOGICAL
MATERIAL
.
43
2.1.7.
SOFTWARE
.
44
2.2.
METHODS
.
45
2.2.1.
YEAST
GROWTH
CONDITIONS
.
45
2.2.2.
TRANSFORMATION
OF
YEAST
.
46
7
2.2.3.
LIBRARY
CONSTRUCTION
AND
VALIDATION
.
47
2.2.4.
ADDITIONAL
YEAST
STRAINS
.
48
2.2.5.
LIBRARY
MEASUREMENT
.
48
2.2.6.
DATA
ANALYSIS
.
49
2.2.7.
PLATE
READER
MEASUREMENTS
.
49
2.2.8.
MICROSCOPY
.
50
2.2.9.
CELL
LYSIS,
SDS-PAGE,
AND
IMMUNOBLOTTING
.
YY
.
.
.
50
2.2.10.
COVALENT
MODIFICATION
WITH
METHOXYPOLYETHYLENEGLYCOL
MALEIMIDE
(PEG-MAL)
.
50
2.2.11.
FLOW
CYTOMETRY
.
51
2.2.12.
METABOLIC
SHIFT
EXPERIMENT
OF
SELECTED
STRAINS
.
51
2.2.13.
HYPOXIA
MEASUREMENTS
.
51
2.2.14.
ASNL-ROGFP2
MICROSCOPY
AND
PLATE
READER
ASSAY
.
52
3.
RESULTS
53
3.1.
LIBRARY
CONSTRUCTION,
MEASUREMENT
AND
ANALYSIS
.
53
3.2.
IDENTIFICATION
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
.
.
63
3.3.
IDENTIFICATION
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
USING
AN
ALTERNATIVE
APPROACH
.
73
3.4.
FUSION
PROTEIN
SPECIFIC
REDOX
DIFFERENCES
ACROSS
DIFFERENT
METABOLIC
CONDITIONS
.
81
3.5.
OXIDATION
OF
ASNL-ROGFP2
CORRELATES
WITH
THE
FORMATION
OF
CYTOOPHIDIAE
.
86
4.
DISCUSSION
93
4.1.
CHALLENGES
OF
C-TERMINAL
TAGGING
IN
GENERAL
.
93
4.2.
CHALLENGES
OF
ENDOGENOUS
C-TERMINAL
TAGGING
WITH
ROGFP2
.
94
4.3.
GENERAL
OBSERVATIONS
.
96
4.4.
LOCAL
AND
CONTEXT
SPECIFIC
ROGFP2
OXIDATION
.
97
4.5.
POSSIBLE
OXIDATION
OF
ROGFP2
ASSOCIATED
PROTEINS
.
100
4.6.
CONCLUSIONS
AND
FUTURE
DIRECTIONS
.
101
5.
REFERENCES
107
8
APPENDICES
121
A.
APPENDIX
123
A.1.
LIST
OF
ROGFP2
FUSION
PROTEINS
FOUND
TO
BE
OXIDISED
USING
THE
R/R
APPROACH
.
123
A.
2.
GO
ENRICHMENT
INCLUDING
SEMANTIC
SIMILARITY
CLUSTERING
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
(R/R
APPROACH)
.
126
A.
3.
LIST
OF
ROGFP2
FUSION
PROTEINS
FOUND
TO
BE
OXIDISED
USING
THE
485/485
APPROACH
.
131
A.
4.
GO
ENRICHMENT
INCLUDING
SEMANTIC
SIMILARITY
CLUSTERING
OF
OXIDISED
ROGFP2
FUSION
PROTEINS
(485/485
APPROACH)
.
148
9 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Bosch, Katharina 1989- |
| author_GND | (DE-588)1278718362 |
| author_facet | Bosch, Katharina 1989- |
| author_role | aut |
| author_sort | Bosch, Katharina 1989- |
| author_variant | k b kb |
| building | Verbundindex |
| bvnumber | BV048918064 |
| ctrlnum | (OCoLC)1372146732 (DE-599)DNB1278052933 |
| discipline | Biologie |
| discipline_str_mv | Biologie |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01546nam a2200373 c 4500</leader><controlfield tag="001">BV048918064</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">230428s2022 gw a||| m||| 00||| eng d</controlfield><datafield tag="015" ind1=" " ind2=" "><subfield code="a">23,H03</subfield><subfield code="2">dnb</subfield></datafield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">1278052933</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="020" ind1=" " ind2=" "><subfield code="c">Broschur</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)1372146732</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)DNB1278052933</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rda</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="044" ind1=" " ind2=" "><subfield code="a">gw</subfield><subfield code="c">XA-DE</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-355</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="8">1\p</subfield><subfield code="a">570</subfield><subfield code="2">23sdnb</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">570</subfield><subfield code="2">sdnb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Bosch, Katharina</subfield><subfield code="d">1989-</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1278718362</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability</subfield><subfield code="c">presented by M.Sc. Katharina Bosch</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Heidelberg</subfield><subfield code="c">2022</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">v, 170 Seiten</subfield><subfield code="b">Illustrationen, Diagramme</subfield><subfield code="c">30 cm</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="b">Dissertation</subfield><subfield code="c">Ruperto Carola University Heidelberg, Germany</subfield><subfield code="d">2022</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">B:DE-101</subfield><subfield code="q">application/pdf</subfield><subfield code="u">https://d-nb.info/1278052933/04</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034182159&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-034182159</subfield></datafield><datafield tag="883" ind1="2" ind2=" "><subfield code="8">1\p</subfield><subfield code="a">dnb</subfield><subfield code="d">20230215</subfield><subfield code="q">DE-101</subfield><subfield code="u">https://d-nb.info/provenance/plan#dnb</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV048918064 |
| illustrated | Illustrated |
| index_date | 2024-07-03T21:55:08Z |
| indexdate | 2024-07-10T09:49:47Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034182159 |
| oclc_num | 1372146732 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | v, 170 Seiten Illustrationen, Diagramme 30 cm |
| publishDate | 2022 |
| publishDateSearch | 2022 |
| publishDateSort | 2022 |
| record_format | marc |
| spelling | Bosch, Katharina 1989- Verfasser (DE-588)1278718362 aut Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability presented by M.Sc. Katharina Bosch Heidelberg 2022 v, 170 Seiten Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Ruperto Carola University Heidelberg, Germany 2022 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1278052933/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034182159&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p dnb 20230215 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Bosch, Katharina 1989- Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |
| subject_GND | (DE-588)4113937-9 |
| title | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |
| title_auth | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |
| title_exact_search | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |
| title_exact_search_txtP | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |
| title_full | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability presented by M.Sc. Katharina Bosch |
| title_fullStr | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability presented by M.Sc. Katharina Bosch |
| title_full_unstemmed | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability presented by M.Sc. Katharina Bosch |
| title_short | Proteome-wide roGFP2-tagging reveals protein context-specific thiol oxidation and its modulation by nutrient availability |
| title_sort | proteome wide rogfp2 tagging reveals protein context specific thiol oxidation and its modulation by nutrient availability |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1278052933/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034182159&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT boschkatharina proteomewiderogfp2taggingrevealsproteincontextspecificthioloxidationanditsmodulationbynutrientavailability |
Es ist kein Print-Exemplar vorhanden.
Inhaltsverzeichnis