Membrane topology and processing of the minor glycoproteins of equine arteritis virus:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
Mensch und Buch Verl.
2013 [erschienen] 2014
|
| Schlagworte: | |
| Online-Zugang: | kostenfrei Inhaltsverzeichnis |
| Beschreibung: | Nebentitel: Membrantopologie und Prozessierung der Glykoproteine Gp2/3/4 des Equinen Arteritis Virus |
| Beschreibung: | 109 S. Ill., graph. Darst. |
| ISBN: | 9783863874735 |
Internformat
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| 245 | 1 | 0 | |a Membrane topology and processing of the minor glycoproteins of equine arteritis virus |c vorgelegt von Anna Karolina Matczuk |
| 246 | 1 | 3 | |a Membrantopologie und Prozessierung der Glykoproteine Gp2/3/4 des Equinen Arteritis Virus |
| 264 | 1 | |a Berlin |b Mensch und Buch Verl. |c 2013 [erschienen] 2014 | |
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| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Nebentitel: Membrantopologie und Prozessierung der Glykoproteine Gp2/3/4 des Equinen Arteritis Virus | ||
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| 650 | 7 | |a Equine arteritis virus |2 cabt | |
| 650 | 7 | |a glycoproteins |2 cabt | |
| 650 | 7 | |a membranes |2 cabt | |
| 650 | 7 | |a signal peptide |2 cabt | |
| 650 | 7 | |a endoplasmic reticulum |2 cabt | |
| 650 | 7 | |a oligosaccharides |2 cabt | |
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Datensatz im Suchindex
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| adam_text | TABIC OF CONTENTS
TABLE OF CONTENTS 1
LIST OF FIGURES 4
LIST OF TABLES 6
ABBREVIATIONS 7
CHAPTER 1 10
INTRODUCTION 10
1.1 TAXONOMY 10
1.2 STRUCTURE OF THE VIRION 10
1.3 TROPISM AND REPLICATION CYCLE 11
1.4 ENVELOPE PROTEINS OF EAV 13
1.5 TARGETING MEMBRANE PROTEINS TO ER 16
1.6 PROTEIN MODIFICATION IN ER: N-LINKED GLYCOSYLATION AND DISULPHIDE
BOND FORMATION. 19
1.6 TRAFFICKING AND RETENTION SIGNALS IN ER 20
1.7 AIMS OF THE STUDY 21
1.8 REFERENCES 22
CHAPTER 2 26
CO-TRANSLATIONAL PROCESSING OF GLYCOPROTEIN 3 FROM EQUINE ARTERITIS
VIRUS: 26
N-GLYCOSYLATION ADJACENT TO THE SIGNAL PEPTIDE PREVENTS CLEAVAGE 26
2.1 ABSTRACT 26
2.2 INTRODUCTION 26
2.3 EXPERIMENTAL PROCEDURES 28
2.3.1 CLONING AND TRANSFECTION 28
2.3.2 METABOLIC LABELING, TUNICAMYCIN TREATMENT, IMMUNOPRECIPITATION AND
FLUOROGRAPHY 29
2.3.3 GLYCOSIDASE TREATMENT 29
2.3.4 IN VITRO TRANSCRIPTION/TRANSLATION 30
2.3.5 PREPARATION OF MEMBRANES AND SOLUBLE FRACTIONS FROM MICROSOMES 30
2.3.6 EXPERIMENTS TO ANALYZE SECRETION OF GP3 FROM CELLS 30
2.3.7 SDS-PAGE, WESTERN BLOTTING, FLUOROGRAPHY 31
2.3.8 FLUORESCENCE PROTEASE PROTECTION ASSAY 31
2.3.9 BIOINFORMATIC IDENTIFICATION OF PROTEINS THAT CONTAIN A
GLYCOSYLATION SITE ADJACENT
TO SIGNAL PEPTIDE CLEAVAGE SITE 31
2.4 RESULTS 32
2.4.1 THE OVERLAPPING SEQUON NEAR THE SIGNAL PEPTIDE IS GLYCOSYLATED AT
BOTH
ASPARAGINES 32
2.4.2 REMOVAL OF TWO GLYCOSYLATION SITES ALLOWS CLEAVAGE OF THE SIGNAL
PEPTIDE 36
2.4.4 THE SIGNAL PEPTIDE OF GP3 DOES NOT FUNCTION AS A MEMBRANE ANCHOR
39
2.4.5 BIOINFORMATIC ANALYSES ON THE OCCURRENCE OF GLYCOSYLATION SITES
ADJACENT TO SIGNAL
PEPTIDES 45
1
HTTP://D-NB.INFO/1051840309
2.5 DISCUSSION 47
2.6 ACKNOWLEDGEMENTS 51
2.7 FOOTNOTES 51
2.8 REFERENCES 52
CHAPTER 3 55
SIGNAL PEPTIDE CLEAVAGE FROM GP3 ENABLED BY REMOVAL OF ADJACENT
GLYCOSYLATION SITES DOES
NOT IMPAIR REPLICATION OF EQUINE ARTERITIS VIRUS IN CELL CULTURE 55
3.1 SUMMARY 55
3.2 MANUSCRIPT 55
3.3 ACKNOWLEDGEMENTS 65
3.4 REFERENES 66
CHAPTER 4 69
LOCALISATION OF THE GP2/4 COMPLEX OF EAV WITH EXCHANGED POTENTIAL
RETENTION SIGNALS AND
THE EFFECT OF SINGLE CYSTEINE MUTATIONS ON THE COMPLEX FORMATION 69
4.1 ABSTRACT 69
4.2 INTRODUCTION 69
4.3 MATERIALS AND METHODS 72
4.3.1 CELLS 72
4.3.2 PLASMIDS 72
4.3.3 FLUORESCENCE PROTEASE PROTECTION ASSAY 73
4.3.4 METABOLIC LABELLING OF PROTEINS 73
4.3.5 IMMUNOPRECIPITATION AND GLYCOSIDASE TREATMENT 73
4.3.6 SDS-PAGE, WESTERN BLOTTING, FLUOROGRAPHY 74
4.3.7 IMMUNOFLUORESCENCE AND CONFOCAL MICROSCOPY 74
4.3.8 REVERSE GENETICS 74
4.4 RESULTS 75
4.4.1 GP2-YFP AND GP4-YFP SHOW TYPE I MEMBRANE TOPOLOGY AND LOCALISE TO
THE ER75
4.4.2 GP2-CFP AND GP4-YFP DO NOT PASS THE MEDIAL GOLGI COMPARTMENT 76
4.4.5 THE CYSTEINE RESIDUES IN THE ECTODOMAIN OF EAV-GP4 ARE ESSENTIAL
FOR VIRUS
INFECTIVITY 82
4.5 DISCUSSION 83
4.6 REFERENCES 85
CHAPTER 5 88
GENERAL DISCUSSION 88
5.1 MEMBRANE TOPOLOGY OF GP3 88
5.2 THE HYDROPHOBIC C-TERMINUS IS RESPONSIBLE FOR MEMBRANE ASSOCIATION
OF GP3 90
5.3 N-LINKED GLYCANS ADJACENT TO THE SIGNAL PEPTIDE CLEAVAGE SITE IN GP3
INTERFERE WITH
CLEAVAGE 94
5.4 OVERLAPPING NNTT SEQUON OF GP3 IS GLYCOSYLATED AT BOTH ASPARAGINES
96
5.5 THE EFFECT OF N-LINKED GLYCANS ADJACENT THE SP CLEAVAGE SITE OF GP3
ON EAV
INFECTIVITY 97
2
5.6 GP2 AND GP4 MEMBRANE TOPOLOGY, ER RETENTION AND DIMERIZATION 98
5.7 FINAL REMARKS AND OUTLOOK 99
5.8 REFERENCES 100
ZUSAMMENFASSUNG 103
SUMMARY 105
PUBLICATIONS 107
ACKNOWLEDGEMENTS 108
3
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| spelling | Matczuk, Anna Karolina Verfasser (DE-588)1051987849 aut Membrane topology and processing of the minor glycoproteins of equine arteritis virus vorgelegt von Anna Karolina Matczuk Membrantopologie und Prozessierung der Glykoproteine Gp2/3/4 des Equinen Arteritis Virus Berlin Mensch und Buch Verl. 2013 [erschienen] 2014 109 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Nebentitel: Membrantopologie und Prozessierung der Glykoproteine Gp2/3/4 des Equinen Arteritis Virus Berlin, Freie Univ., Diss., 2014 Equine arteritis virus cabt glycoproteins cabt membranes cabt signal peptide cabt endoplasmic reticulum cabt oligosaccharides cabt translocation cabt Glykoproteine (DE-588)4071905-4 gnd rswk-swf Signaltransduktion (DE-588)4318717-1 gnd rswk-swf Membranproteine (DE-588)4130026-9 gnd rswk-swf Equines Arteriitis-Virus (DE-588)4367969-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Equines Arteriitis-Virus (DE-588)4367969-9 s Glykoproteine (DE-588)4071905-4 s Membranproteine (DE-588)4130026-9 s Signaltransduktion (DE-588)4318717-1 s DE-604 Elektronische Reproduktion urn:nbn:de:kobv:188-fudissthesis000000096791-5 http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000096791 kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027341535&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Matczuk, Anna Karolina Membrane topology and processing of the minor glycoproteins of equine arteritis virus Equine arteritis virus cabt glycoproteins cabt membranes cabt signal peptide cabt endoplasmic reticulum cabt oligosaccharides cabt translocation cabt Glykoproteine (DE-588)4071905-4 gnd Signaltransduktion (DE-588)4318717-1 gnd Membranproteine (DE-588)4130026-9 gnd Equines Arteriitis-Virus (DE-588)4367969-9 gnd |
| subject_GND | (DE-588)4071905-4 (DE-588)4318717-1 (DE-588)4130026-9 (DE-588)4367969-9 (DE-588)4113937-9 |
| title | Membrane topology and processing of the minor glycoproteins of equine arteritis virus |
| title_alt | Membrantopologie und Prozessierung der Glykoproteine Gp2/3/4 des Equinen Arteritis Virus |
| title_auth | Membrane topology and processing of the minor glycoproteins of equine arteritis virus |
| title_exact_search | Membrane topology and processing of the minor glycoproteins of equine arteritis virus |
| title_full | Membrane topology and processing of the minor glycoproteins of equine arteritis virus vorgelegt von Anna Karolina Matczuk |
| title_fullStr | Membrane topology and processing of the minor glycoproteins of equine arteritis virus vorgelegt von Anna Karolina Matczuk |
| title_full_unstemmed | Membrane topology and processing of the minor glycoproteins of equine arteritis virus vorgelegt von Anna Karolina Matczuk |
| title_short | Membrane topology and processing of the minor glycoproteins of equine arteritis virus |
| title_sort | membrane topology and processing of the minor glycoproteins of equine arteritis virus |
| topic | Equine arteritis virus cabt glycoproteins cabt membranes cabt signal peptide cabt endoplasmic reticulum cabt oligosaccharides cabt translocation cabt Glykoproteine (DE-588)4071905-4 gnd Signaltransduktion (DE-588)4318717-1 gnd Membranproteine (DE-588)4130026-9 gnd Equines Arteriitis-Virus (DE-588)4367969-9 gnd |
| topic_facet | Equine arteritis virus glycoproteins membranes signal peptide endoplasmic reticulum oligosaccharides translocation Glykoproteine Signaltransduktion Membranproteine Equines Arteriitis-Virus Hochschulschrift |
| url | http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000096791 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027341535&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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