Structural characterisation of protein derived N-glycans by mass spectrometry: application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation
Saved in:
| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
2001
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Münster, Univ., Diss., 2001 |
| Physical Description: | VII, 157 S. Ill., graph. Darst. |
Staff View
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV014314831 | ||
| 003 | DE-604 | ||
| 005 | 20030523 | ||
| 007 | t | ||
| 008 | 020527s2001 ad|| m||| 00||| eng d | ||
| 016 | 7 | |a 964761831 |2 DE-101 | |
| 035 | |a (OCoLC)248974185 | ||
| 035 | |a (DE-599)BVBBV014314831 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-91 |a DE-29T |a DE-355 | ||
| 100 | 1 | |a Šagi, Dijana |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Structural characterisation of protein derived N-glycans by mass spectrometry |b application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation |c vorgelegt von Dijana Šagi |
| 246 | 1 | |a Structural characterisation of protein-derived N-glycans by mass spectrometry | |
| 264 | 1 | |c 2001 | |
| 300 | |a VII, 157 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Münster, Univ., Diss., 2001 | ||
| 650 | 0 | 7 | |a Massenspektrometrie |0 (DE-588)4037882-2 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Glucane |0 (DE-588)4157662-7 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Glucane |0 (DE-588)4157662-7 |D s |
| 689 | 0 | 1 | |a Massenspektrometrie |0 (DE-588)4037882-2 |D s |
| 689 | 0 | |5 DE-604 | |
| 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=009820392&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009820392 | |
Record in the Search Index
| _version_ | 1812460567602921472 |
|---|---|
| adam_text |
CONTENTS
_I
CONTENTS
ABBREVIATIONS.
V
1 INTRODUCTION. 1
1.1 MASS SPECTROMETRY IN ANALYTICAL BIOCHEMISTRY. 1
1.1.1 IMPORTANT TERMS IN MASS SPECTROMETRY. 1
1.1.2 INSTRUMENTATION. 2
1.1.2.1 ION SOURCE. 3
1.1.2.1.1 MATRIX-ASSISTED LASER DESORPTION/IONISATION (MALDI). 3
1.1.2.1.2 ELECTROSPRAY IONISATION (ESI). 4
1.1.2.2 MASS ANALYSERS.-. 7
1.1.2.2.1 TIME-OF-FLIGHT (TOF). 7
1.1.2.2.2 QUADRUPOLE MASS ANALYSER. 10
1.1.2.3 TANDEM MASS SPECTROMETRY AND CID. 13
1.2 N-GLYCOSYLATIONS OF PROTEINS.:. 14
1.2.1 BIOSYNTHESES OF N-LINKED OLIGOSACCHARIDES. 15
1.2.2 FUNCTION OF N-LINKED OLIGOSACCHARIDES. 19
1.2.3 E-SELECTIN AND E-SELECTIN LIGAND-1. 20
1.2.4 CONGENITAL DISORDERS OF GLYCOSYLATION (CDG). 24
1.2.4.1 DIAGNOSIS OF CDG-TYPES. 25
1.2.5 STRUCTURAL ANALYSIS QF N-GLYCANS. 27
1.2.5.1 DETECTION OF N-GLYCOSYLATION. 27
1.2.5.2 RELEASE OF N-GLYCANS. 27
1.2.5.3 CHROMATOGRAPHIC PROFILING OF RELEASED N-GLYCANS. 28
1.2.5.4 MONOSACCHARIDE COMPOSITION ANALYSIS.
/.
. 30
1.2.5.5 METHYLATION ANALYSIS. 30
1.2.5.6 NUCLEAR MAGNETIC RESONANCE (NMR). 31
1.2.5.7 SEQUENTIAL EXOGLYCOSIDASE DIGESTION. 31
1.2.5.8 MASS SPECTROMETRY. 31
2 OBJECTIVES OF THE STUDY.
35
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/964761831
CONTENTS
_N
3 EXPERIMENTAL PART. 37
3.1 IN-SOLUTION DEGLYCOSYLATION. 37
3.2 IN-GEL DEGLYCOSYLATION. 38
3.2.1 SODIUMDODECYLSULPHATE-POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE). 38
3.2.2 PROTEIN REDUCTION AND ALKYLATION. 39
3.2.3 ENZYMATIC RELEASE OF N-GLYCANS IN-GEL. 39
3.2.4 EXTRACTION OF N-GLYCANS AFTER IN-GEL DEGLYCOSYLATION. 39
3.3 DESALTING OF NATIVE N-GLYCANS. 40
3.4 METHYLATION OF SIALIC ACIDS. 40
3.5 DESALTING OF METHYLATED N-GLYCANS. 40
3.6 ENZYMATIC REMOVAL OF SIALIC ACIDS. 41
3.7 IN-GEL TRYPTIC DIGESTION. 41
3.7.1 PEPTIDE EXTRACTION AND PURIFICATION. 41
3.8 MASS SPECTROMETRY. 42
3.8.1 MALDITOF MS. 42
3.8.2 ESI Q-TOF MS. 45
3.9 HPAEC/PAD. 47
3.10 STANDARD N-GLYCANS. 49
4 RESULTS AND DISCUSSION. 51
4.1 METHODS FOR STRUCTURAL ANALYSIS AND SEQUENCING OF N-GLYCANS. 51
4.1.1 N-DEGLYCOSYLATION OF PROTEINS. 51
4.1.1.1 IN-SOLUTION DEGLYCOSYLATION OF PROTEIN MIXTURE. 52
4.1.1.2 IN-GEL DEGLYCOSYLATION OF PROTEIN COMPONENTS FROM THE
MIXTURES. 52
4.1.2 PURIFICATION OF THE RELEASED N-GLYCANS. 53
4.1.3 METHYLATION OF SIALIC ACIDS. 55
4.1.4 MS ANALYSIS. 58
4.1.4.1 N-GLYCANS PROFILING BY MALDI MS. 58
4.1.4.1.1 NEUTRAL N-GLYCANS. 58
4.1.4.1.2 SIALYLATED N-GLYCANS AND MIXTURES OF SIALYLATED AND
NEUTRAL N-GLYCANS. 59
4.1.4.1.3 METHYLATED SIALYLATED N-GLYCANS. 63
CONTENTS
4.1.4.1.4 QUANTITATIVE ASPECTS OF MALDI MS OF N-GLYCANS. 63
4.1.4.2 N-GLYCAN ANALYSIS BY ESI Q-TOF MS. 59
4.1.4.2.1 SEQUENCING OF NEUTRAL N-GLYCANS BY (+)ESI Q-TOF
MS. 65
4.1.4.2.2 SEQUENCING OF SIALYLATED N-GLYCANS (-)ESI Q-TOF
MS.*. 72
4.2 STRUCTURAL DETERMINATION OF N-GLYCANS OF E-SELECTIN LIGAND-1
GLYCOPROTEIN. 82
4.2.1 ANALYSIS OF ESL-1 MYC PROTEIN EXPRESSED IN CHO-PRO'5 CELLS. 82
4.2.1.1 IDENTIFICATION OF THE PROTEIN SPECIES IN THE ADDITIONAL MAJOR
BAND IN SDS-PAGE. 84
4.2.1.2 ANALYSIS OF N-GLYCANS AFTER IN-SOLUTION DEGLYCOSYLATION. 84
4.2.1.2.1 NATIVE N-GLYCAN FIXTURE.,T. 85
4.2.1.2.2 NEUTRAL AND METHYLATED SIALYLATED N-GLYCAN MIXTURE. 90
4.2.1.3 ANALYSIS OF N-GLYCANS AFTER IN-GEL DEGLYCOSYLATION. 92
4.2.2 ANALYSIS OF ESL-1 MYC PROTEIN EXPRESSED IN CHO-DHFR' CELLS. 94
4.2.2.1 ANALYSIS OF N-GLYCANS AFTER IN-SOLUTION DEGLYCOSYLATION. 94
4.2.2.1.1 NATIVE N-GLYCAN MIXTURE. 94
4.2.2.1.2 NEUTRAL AND METHYLATED SIALYLATED N-GLYCAN MIXTURE. 98
4.2.2.2 ANALYSIS OF N-GLYCANS AFTER IN-GEL DEGLYCOSYLATION.100
4.2.2.3 ESI MS/MS ANALYSIS.102
4.3 ABERRANT N-GLYCOSYLATION IN CONGENITAL DISORDERS OF
GLYCOSYLATION.106
4.3.1 CLINICAL PHENOTYPE OF THE CDG PATIENT.106
4.3.2 ISOELECTRIC FOCUSING OF SERUM TRANSFERRIN.107
4.3.3 N-GLYCOSYLATION OF SERUM TRANSFERRIN.107
4.3.4 N-GLYCOSYLATION OF SERUM AL-ANTITRYPSIN.114
4.3.5 N-GLYCOSYLATION OF PLASMA PROTEINS.
T.116
4.3.5.1 ANALYSIS OF MIXTURE OF NEUTRAL AND SIALYLATED N-GLYCANS.116
4.3.5.1.1 MALDI MS.117
4.3.5.1.2 HPAEC/PAD.118
4.3.5.2 ANALYSIS OF DESIALYLATED N-GLYCANS.123
4.3.5.2.1 MALDI MS.123
4.3.5.2.2 HPAEC/PAD. 125
133
5 SUMMARY.
CONTENTS
IV
6 REFERENCES. 139
7 ACKNOWLEDGEMENTS. 153
8 PUBLICATIONS AND PRESENTATIONS. 155
9 LEBENSLAUF. 157 |
| any_adam_object | 1 |
| author | Šagi, Dijana |
| author_facet | Šagi, Dijana |
| author_role | aut |
| author_sort | Šagi, Dijana |
| author_variant | d š dš |
| building | Verbundindex |
| bvnumber | BV014314831 |
| ctrlnum | (OCoLC)248974185 (DE-599)BVBBV014314831 |
| format | Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>00000nam a2200000 c 4500</leader><controlfield tag="001">BV014314831</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20030523</controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">020527s2001 ad|| m||| 00||| eng d</controlfield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">964761831</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)248974185</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BVBBV014314831</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-91</subfield><subfield code="a">DE-29T</subfield><subfield code="a">DE-355</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Šagi, Dijana</subfield><subfield code="e">Verfasser</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Structural characterisation of protein derived N-glycans by mass spectrometry</subfield><subfield code="b">application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation</subfield><subfield code="c">vorgelegt von Dijana Šagi</subfield></datafield><datafield tag="246" ind1="1" ind2=" "><subfield code="a">Structural characterisation of protein-derived N-glycans by mass spectrometry</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2001</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">VII, 157 S.</subfield><subfield code="b">Ill., graph. Darst.</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="500" ind1=" " ind2=" "><subfield code="a">Münster, Univ., Diss., 2001</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Massenspektrometrie</subfield><subfield code="0">(DE-588)4037882-2</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Glucane</subfield><subfield code="0">(DE-588)4157662-7</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</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="689" ind1="0" ind2="0"><subfield code="a">Glucane</subfield><subfield code="0">(DE-588)4157662-7</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="1"><subfield code="a">Massenspektrometrie</subfield><subfield code="0">(DE-588)4037882-2</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2=" "><subfield code="5">DE-604</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=009820392&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="943" ind1="1" ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-009820392</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV014314831 |
| illustrated | Illustrated |
| indexdate | 2024-10-09T18:04:50Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009820392 |
| oclc_num | 248974185 |
| open_access_boolean | |
| owner | DE-91 DE-BY-TUM DE-29T DE-355 DE-BY-UBR |
| owner_facet | DE-91 DE-BY-TUM DE-29T DE-355 DE-BY-UBR |
| physical | VII, 157 S. Ill., graph. Darst. |
| publishDate | 2001 |
| publishDateSearch | 2001 |
| publishDateSort | 2001 |
| record_format | marc |
| spelling | Šagi, Dijana Verfasser aut Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation vorgelegt von Dijana Šagi Structural characterisation of protein-derived N-glycans by mass spectrometry 2001 VII, 157 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Münster, Univ., Diss., 2001 Massenspektrometrie (DE-588)4037882-2 gnd rswk-swf Glucane (DE-588)4157662-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Glucane (DE-588)4157662-7 s Massenspektrometrie (DE-588)4037882-2 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009820392&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Šagi, Dijana Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation Massenspektrometrie (DE-588)4037882-2 gnd Glucane (DE-588)4157662-7 gnd |
| subject_GND | (DE-588)4037882-2 (DE-588)4157662-7 (DE-588)4113937-9 |
| title | Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation |
| title_alt | Structural characterisation of protein-derived N-glycans by mass spectrometry |
| title_auth | Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation |
| title_exact_search | Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation |
| title_full | Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation vorgelegt von Dijana Šagi |
| title_fullStr | Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation vorgelegt von Dijana Šagi |
| title_full_unstemmed | Structural characterisation of protein derived N-glycans by mass spectrometry application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation vorgelegt von Dijana Šagi |
| title_short | Structural characterisation of protein derived N-glycans by mass spectrometry |
| title_sort | structural characterisation of protein derived n glycans by mass spectrometry application to glycosylation analysis of e selectin ligand 1 and potentials in diagnosis of congenital disorders of glycosylation |
| title_sub | application to glycosylation analysis of E-selectin ligand-1 and potentials in diagnosis of congenital disorders of glycosylation |
| topic | Massenspektrometrie (DE-588)4037882-2 gnd Glucane (DE-588)4157662-7 gnd |
| topic_facet | Massenspektrometrie Glucane Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009820392&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT sagidijana structuralcharacterisationofproteinderivednglycansbymassspectrometryapplicationtoglycosylationanalysisofeselectinligand1andpotentialsindiagnosisofcongenitaldisordersofglycosylation AT sagidijana structuralcharacterisationofproteinderivednglycansbymassspectrometry |