Post-translational processing a practical approach
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| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford [u.a.]
Oxford Univ. Press
1999
|
| Schriftenreihe: | The practical approach series
203 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
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| 245 | 1 | 0 | |a Post-translational processing |b a practical approach |c ed. by S. J. Higgins ... |
| 264 | 1 | |a Oxford [u.a.] |b Oxford Univ. Press |c 1999 | |
| 300 | |a XX, 317 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 1 | |a The practical approach series |v 203 | |
| 650 | 7 | |a Protéines - Modifications posttraductionnelles |2 ram | |
| 650 | 4 | |a Post-translational modification | |
| 700 | 1 | |a Higgins, Stephen J. |e Sonstige |4 oth | |
| 830 | 0 | |a The practical approach series |v 203 |w (DE-604)BV007921321 |9 203 | |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-008675814 | |
Datensatz im Suchindex
| DE-19_call_number | 1900/WD 5100 H636 |
|---|---|
| DE-19_location | 29 |
| DE-BY-UBM_katkey | 1844255 |
| DE-BY-UBM_media_number | 41619851750014 |
| _version_ | 1823051565181698048 |
| adam_text | Contents
List of Con trib u tors xv
Abbreviations xvii
1. Sequence analysis of expressed proteins 1
JeffN. Keen and Alison E. Ashcroft
1. Introduction 1
2. N terminal sequence analysis 3
Automated sequencing 3
3. Sample preparation 9
Sample preparation by SDS PAGE 10
Sample preparation by HPLC 12
Other procedures for sample preparation 12
4. N terminal blocking 14
Protein fragmentation 17
5. C terminal sequencing 21
Chemical analysis of the C terminus 21
Enzymic analysis of the C terminus 22
6. Mass spectrometric analysis of proteins 25
A brief guide to mass spectrometers 25
Molecular mass determination 29
Sequencing by mass spectrometry 34
References 40
2. Protein folding and import into organelles 43
Joachim Rassow
1. Introduction 43
2. Preparation of preprotein substrates for in vitro import into
organelles 44
Preparation of mRNA for in vitro translation 44
Synthesis of preproteins in the reticulocyte lysate 44
Synthesis of preproteins in wheat germ lysate 46
Synthesis of preproteins in yeast cytosol 47
Synthesis of preproteins in Escherichia coli 47
Dihydrofolate reductase as a model protein for import studies 47
Contents
3. Import of proteins into mitochondria 49
Isolation of mitochondria for import studies 50
Import of preproteins into isolated mitochondria 56
Generation of translocation intermediates 63
Localization of imported proteins 68
4. Import of proteins into microsomes (endoplasmic reticulum) 70
Isolation of microsomes 70
Import of preproteins into microsomes 73
5. Import of proteins into other organelles 74
6. Analysis of protein import into organelles 75
Monitoring the association of proteins with membranes 75
Analysis of protein complexes involved in organelle import 77
Analysis of protein folding after import into organelles 90
Acknowledgements 91
References 92
3. Analysis of protein phosphorylation 95
5. Ivar Walaas and Anne Carine 0stvold
1. Introduction 95
2. Investigating protein phosphorylation systems 96
3. Phosphorylation of proteins in intact preparations 96
General considerations 96
Intact animals 96
Tissue slices 97
Isolated cells 98
4. Phosphorylation of proteins in cell free preparations 99
General considerations 99
Labelling and stimulation of cell free preparations 100
5. Analysis of phosphorylated proteins 101
General considerations 101
Quantification of phosphoproteins 101
Phosphoprotein separation 102
Protein isolation by immunomethods 103
Analysis of multisite phosphorylation 103
Phosphoamino acid analysis 107
Analysis of the state of phosphorylation 108
6. Analysis of protein kinases 110
General considerations 110
Analysis of protein kinase activity in vitro 110
Analysis of specific protein kinases in vitro 113
x
Contents
Analysis of protein kinase activity in intact cells 122
Purification of protein kinases 123
7. Phosphoprotein phosphatases 123
General considerations 123
P Ser/P Thr phosphoprotein phosphatases 123
Tyrosine specific phosphoprotein phosphatases 127
References 128
4. Protein glycosylation 135
David A. Ashford and Fran Platt
1. Introduction 135
Protein glycosylation 135
Oligosaccharide structures 136
Glycosylation pathways 137
Characterization of protein glycosylation 140
2. Is my protein glycosylated? 140
Colorimetric methods 140
Proprietary detection methods 141
Lectin binding 142
3. Study of whole protein glycosylation 144
Detection of/V glycosylation 144
Monosaccharide composition 146
Lectin binding analysis 149
Susceptibility to endoglycosidase H 150
Other methods 151
4. Analysis of glycosylation sites 152
Protease mapping of glycopeptides 152
Glycopeptide identification and analysis 154
5. Analysis of glycan structure 155
Glycan release 155
Glycan labelling 159
Glycan separation 161
Structural analysis of glycans 167
6. Manipulation of protein glycosylation 170
References 172
5. Lipid modification of proteins 175
Nigel M. Hooper and R. A. Jeffrey Mcllhinney
1. Introduction 175
xi
Contents
2. Protein acylation 175
General considerations 175
Enzymology 176
3. Identification of acylated proteins 177
General points 177
Labelling cultured cells with fatty acids 179
Analysis of acylated proteins 180
Myristoylated proteins 184
4. Glycosyl phosphatidylinositol membrane anchorage of
proteins 188
Structure of GPI anchors 188
Signals for attachment of a GPI anchor to a protein 188
5. Identification of GPI anchorage 190
General points 190
Release of GPI anchored proteins by bacterial
phosphatidylinositol specific phospholipase C 191
Differential detergent solubilization 194
Detection of the cross reacting determinant 195
Metabolic labelling 200
6. Prenylation 200
References 201
6. Proteolytic processing 205
John M. W. Creemers, Elaine M. Bailyes, Iris Lindberg,
and John C. Hutton
1. Introduction 205
2. Immunoadsorbent assay of PCI and PC2 208
Calcium dependence of prohormone convertases 212
3. Expression of prohormone convertases in DG44 CHO cells 213
4. Fluorogenic assay for PCI and PC2 216
PCI assay 217
PC2 assay 218
5. Transient expression with recombinant vaccinia virus V.V.:T7 220
Introduction 220
References 222
7. Protein degradation in mammalian cells 225
Aaron Ciechanover and Bernd Wiederanders
1. The ubiquitin proteasome pathway in mammalian cells 225
xii
Contents
Introduction 225
Preparation of cell extracts for monitoring conjugation and degradation 229
Fractionation of cell extracts for monitoring conjugation and
degradation 230
Labelling of proteolytic substrates 233
Conjugation of proteolytic substrates 236
Degradation of proteolytic substrates 241
Use of inhibitors to study proteasome function 241
2. Proteolysis in mammalian lysosomns 244
Introduction 244
Isolation of mammalian lysosomes 245
Lysosomal peptidases 251
Measurement of lysosomal protein degradation 257
Acknowledgements 261
Roforencos 261
8. Protein degradation and proteinases in yeast 265
Wolfgang Hilt and Dieter //. Wolf
1. Introduction 265
Advantages of yeast for studying cukaryotic cell biology 266
2. Growth of yeast colls and preparation of cell extracts 26X
3. Analysis of protein turnover 270
Protein degradation in vivo 270
Degradation of individual proteins 272
4. The proteasome and protein degradation in yeast 275
5. Proteinase yscD. a major cytoplasmic peptidase 2S3
Introduction 2X3
Assay of proteinase yscD 2X4
6. Protein degradation in the vacuole 2X5
Introduction 2X5
Assay of vacuolar peptidases 2X6
Purification of yeast vacuolar proteinases 2^4
Isolation of yeast vacuoles 2W
Acknowledgements 300
References 301
Appendix 303
Index 3in
xiii
|
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| id | DE-604.BV012759273 |
| illustrated | Illustrated |
| indexdate | 2025-02-03T16:34:53Z |
| institution | BVB |
| isbn | 0199637946 0199637954 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008675814 |
| oclc_num | 40489139 |
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| physical | XX, 317 S. Ill., graph. Darst. |
| publishDate | 1999 |
| publishDateSearch | 1999 |
| publishDateSort | 1999 |
| publisher | Oxford Univ. Press |
| record_format | marc |
| series | The practical approach series |
| series2 | The practical approach series |
| spellingShingle | Post-translational processing a practical approach The practical approach series Protéines - Modifications posttraductionnelles ram Post-translational modification |
| title | Post-translational processing a practical approach |
| title_auth | Post-translational processing a practical approach |
| title_exact_search | Post-translational processing a practical approach |
| title_full | Post-translational processing a practical approach ed. by S. J. Higgins ... |
| title_fullStr | Post-translational processing a practical approach ed. by S. J. Higgins ... |
| title_full_unstemmed | Post-translational processing a practical approach ed. by S. J. Higgins ... |
| title_short | Post-translational processing |
| title_sort | post translational processing a practical approach |
| title_sub | a practical approach |
| topic | Protéines - Modifications posttraductionnelles ram Post-translational modification |
| topic_facet | Protéines - Modifications posttraductionnelles Post-translational modification |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008675814&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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| work_keys_str_mv | AT higginsstephenj posttranslationalprocessingapracticalapproach |