Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
Köster
2014
|
| Schlagworte: | |
| Online-Zugang: | Inhaltstext Inhaltsverzeichnis |
| Beschreibung: | XI, 122 S. Ill., graph. Darst. |
| ISBN: | 9783895748578 |
Internformat
MARC
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| 245 | 1 | 0 | |a Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry |c Ahmed H. El-Khatib |
| 264 | 1 | |a Berlin |b Köster |c 2014 | |
| 300 | |a XI, 122 S. |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
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INDEX
ACKNOWLEDGEMENT I
SUMMARY II
ZUSSAMENFASSUNG V
INDEX VIII
1 INTRODUCTION 1
1.1 PROTEOMICS: AN OVERVIEW 1
1.1.1 PROTEOME 1
1.1.2 SIGNIFICANCE OF POST-TRANSLATIONAL MODIFICATION (PTM) 3
SULFENATION AS PTM 4
1.2 IONIZATION TECHNIQUES IN MASS SPECTROMETRY FOR PROTEOMICS 6
1.2.1 ESI (ELECTROSPRAY IONIZATION) 6
1.2.2 NS1 (NANOSPRAY IONIZATION) 7
1.2.3 MALDI (MATRIX ASSISTED LASER DESORPTION IONIZATION) 8
1.2.4 ICP (INDUCTIVELY COUPLED PLASMA) 8
1.3 FRAGMENTATION OF PEPTIDES AND PROTEINS USING MASS SPECTROMETRY 10
1.3.1 CID (COLLISION-INDUCED DISSOCIATION) 11
1.3.2 IRMPD (INFRARED MULTIPHOTON DISSOCIATION) 11
1.3.3 HCD (HIGHER ENERGY COLLISIONAL DISSOCIATION) 12
1.4 QUANTIFICATION OF PEPTIDES AND PROTEINS USING MASS SPECTROMETRY 12
1.4.1 LABEL-FREE QUANTITATIVE PROTEOMICS 15
1.4.2 QUANTITATIVE PROTEOMICS USING STABLE ISOTOPE LABELING 15
METABOLIC LABELING 15
ENZYMATIC LABELING 17
CHEMICAL LABELING 17
1.4.3 QUANTITATIVE PROTEOMICS USING BI-FUNCTIONAL CHELATING AGENTS AS
CHEMICAL LABELS
AND LANTHANIDES AS MASS MARKERS 23
ECAT (ELEMENT CODED AFFINITY TAG) 24
MECT (METAL ELEMENT CHELATE TAG) 25
1.5 QUANTITATIVE PROTEOMICS VIA NATURAL ELEMENT TAGS AND CHEMICAL LABELS
USING
INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY (ICP-MS) 25
1.5.1 VIA NATURALLY OCCURRING HETEROATOM TAGS 26
SULFUR 26
PHOSPHORUS 27
SELENIUM 27
1.5.2 VIA "ARTIFICIAL" ELEMENTAL LABELING 28
IODINE 28
IRON 28
MERCURY 29
VIII
HTTP://D-NB.INFO/1051044332
MECAT (METAL CODED AFFINITY TAG) USING LANTHANIDES 29
1.6 MULTI-ELEMENTAL LABELING OF PEPTIDES AND PROTEINS 31
POLYMER-BASED ELEMENTAL TAGS 31
NANOPARTICLES 32
2 AIM AND SCOPE OF WORK 34
3 RESULTS AND DISCUSSION 35
3.1 DUAL LABELING OF PEPTIDES USING MECAT AND DOT A DERIVATIVES 35
ADVANTAGES OF CYSTEINYL-PEPTIDE ENRICHMENT ON THIOPROPYL SEPHAROSE 6B
RESIN 38
OPTIMIZATION OF CAPTURE/WASH/RELEASE OF CYSTEINYL-PEPTIDE FROM
THIOPROPYL SEPHAROSE
6B RESIN 39
OPTIMIZATION OF ON-RESIN AMINO LABELING 40
EVALUATION OF ON-RESIN AMINO LABELING 42
PEPTIDE ELUTION FROM THE RESIN 46
EFFECT OF EXCESS TCEP ON MECAT-LA LABELING 47
REMOVAL OF EXCESS TCEP 48
ESTIMATION OF LOSSES 49
THIOL LABELING AND DEMONSTRATION OF DUAL LABELING 50
METHODOLOGICAL SCOPE FOR QUANTITATIVE APPLICATIONS USING STANDARD
ADDITION TECHNIQUE
57
CONCLUSION AND OUTLOOK 58
3.2 LABELING OF PROTEINS AND PEPTIDES USING GOLD 60
MONOMALEIMIDO-UNDECAGOLD 60
LABELING OF PROTEIN WITH MONOMALEIMIDO-UNDECAGOLD 61
C3 LC-ES1-FTICR-MS AND LC-ICP-MS ANALYSIS OF PROTEINS LABELED WITH
MONOMALEIMIDO-UNDECAGOLD 62
SDS-PAGE SEPARATION OF PROTEINS LABELED WITH MONOMALEIMIDO-UNDECAGOLD 62
OFFGEL ELECTROPHORESIS OF PROTEINS LABELED WITH MONOMALEIMIDO-UNDECAGOLD
63
LABELING OF PEPTIDES WITH MONOMALEIMIDO-UNDECAGOLD 65
CONCLUSION AND OUTLOOK 67
3.3 QUANTIFICATION OF SULFENIC ACID MODIFICATIONS IN PEPTIDES AND
PROTEINS USING
MOLECULAR AND ELEMENTAL MASS SPECTROMETRY 68
SYNTHESIS OF LN-DOTA-DIMEDONE 71
PURIFICATION OF LN-DOTA-DIMEDONE 73
IN VITRO OXIDATION OF CYSTEINE RESIDUES AND REACTIVITY WITH DIMEDONE 76
IN VITRO OXIDATION OF PEPTIDES AND LABELING WITH LN-DOTA-DIMEDONE 78
PARALLEL LABELING OF PEPTIDES WITH LN-DOTA-DIMEDONE AND LN-MECAT-IA 82
CONSECUTIVE LABELING OF PEPTIDES WITH LN-DOTA-DIMEDONE AND LN-MECAT-IA
83
OPTIMIZATION OF IN VITRO OXIDATION OF PROTEINS AND LABELING WITH
LN-DOTA-DIMEDONE
84
PARALLEL LABELING OF PROTEINS WITH LN-DOTA-DIMEDONE AND LN-MECAT-IA 85
CONSECUTIVE LABELING OF PROTEINS WITH LN-DOTA-DIMEDONE AND LN-MECAT-IA
86
ESI-MS/MS BASED RELATIVE QUANTIFICATION OF SULFENIC ACID MODIFICATIONS
87
IX
CONCLUSION AND OUTLOOK 90
4 MATERIALS AND METHODS 92
4.1 DUAL LABELING OF PEPTIDES USING MECAT AND DOTA DERIVATIVES 92
CHEMICALS AND MATERIALS 92
SYNTHESIS OF LN-MCCAT-LA 92
PROTEINS AND PEPTIDES 93
SELECTION OF PEPTIDES 93
PROTEIN DIGESTION 94
CYSTEINYL-PEPTIDC ENRICHMENT ON THE THIOPROPYL SEPHAROSE 6B RESIN 94
ON-RCSIN AMINO LABELING 94
PEPTIDE ELUTION FROM THE RESIN AND REMOVAL OF EXCESS TCEP 95
CYSTEINE LABELING WITH LN-MECAT-LA 95
CI8 NANOLC-ESL-FTICR-MS ANALYSIS OF LABELED PEPTIDES 95
C|
8
LC-ESI-FT1CR-MS AND LC-ICP-MS OF LABELED PEPTIDES 96
4.2 LABELING OF PROTEINS AND PEPTIDES USING GOLD 97
CHEMICALS AND MATERIALS 97
PROTEINS AND PEPTIDES 97
LABELING OF PROTEIN WITH MONOMALEIMIDO-UNDECAGOLD 97
C3 LC-ES1-FTICR-MS AND LC-ICP-MS ANALYSIS OF PROTEINS LABELED WITH
MONOMALEIMIDO-UNDECAGOLD 97
SDS-PAGE SEPARATION OF PROTEINS LABELED WITH MONOMALEIMIDO-UNDECAGOLD 98
OFFGEL ELECTROPHORESIS OF PROTEINS LABELED WITH MONOMALEIMIDO-UNDECAGOLD
98
ICP-MS ANALYSIS OF OFFGEL FRACTIONS 99
LABELING OF PEPTIDES WITH MONOMALEIMIDO-UNDECAGOLD 99
CIG LC-ESI-FT1CR-MS AND LC-ICP-MS ANALYSIS OF PEPTIDES LABELED WITH
MONOMALEIMIDO-UNDECAGOLD 99
4.3 QUANTIFICATION OF SULFENIC ACID MODIFICATIONS IN PEPTIDES AND
PROTEINS USING
MOLECULAR AND ELEMENTAL MASS SPECTROMETRY 100
CHEMICALS AND MATERIALS 100
SYNTHESIS OF LN-DOTA-DIMEDONE 100
LC-U V-MS ANALYSIS OF THE SYNTHESIZED LN-DOT A-DIMEDONE 101
PURIFICATION OF THE SYNTHESIZED LN-DOTA-DIMEDONE 101
PROTEINS AND PEPTIDES 101
IN VITRO OXIDATION OF PEPTIDES AND LABELING WITH DIMEDONE 101
IN VITRO OXIDATION OF PEPTIDES AND LABELING WITH LN-DOTA-DIMEDONE 101
LABELING OF PEPTIDES WITH LN-MECAT-LA 102
CIS NANOLC-ESI-FTICR-MS ANALYSIS OF PEPTIDES LABELED WITH DIMEDONE,
LN-DOTA-
DIMEDONE OR LN-MECAT-IA 102
SCX LC-ICP-MS ANALYSIS OF PI LABELED WITH TB-DOTA-DIMEDONE OR
HO-MECAT-LA
102
OXIDATION OF PROTEINS AND LABELING WITH LN-DOTA-DIMEDONE 103
LABELING OF PROTEINS WITH LN-MECAT-IA 103
SDS-PAGE SEPARATION OF PROTEINS 103
X
1CP-MS ANALYSIS OF PROTEIN BANDS IN SDS-PAGE 103
PARALLEL LABELING OF PEPTIDES AND PROTEINS 104
CONSECUTIVE LABELING OF PEPTIDES AND PROTEINS 104
INSTRUMENTAL PARAMETERS FOR IRMPD AND HCD FRAGMENTATION 104
5 REFERENCES 106
6 ABBREVIATIONS 118
LEBENSLAUF 120
PUBLIKATIONSLISTE 122
XI |
| any_adam_object | 1 |
| author | El-Khatib, Ahmed H. 1979- |
| author_GND | (DE-588)1052064426 |
| author_facet | El-Khatib, Ahmed H. 1979- |
| author_role | aut |
| author_sort | El-Khatib, Ahmed H. 1979- |
| author_variant | a h e k ahe ahek |
| building | Verbundindex |
| bvnumber | BV041936129 |
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| ctrlnum | (OCoLC)880704872 (DE-599)DNB1051044332 |
| dewey-full | 540 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 540 - Chemistry and allied sciences |
| dewey-raw | 540 |
| dewey-search | 540 |
| dewey-sort | 3540 |
| dewey-tens | 540 - Chemistry and allied sciences |
| discipline | Chemie / Pharmazie |
| format | Thesis Book |
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| spelling | El-Khatib, Ahmed H. 1979- Verfasser (DE-588)1052064426 aut Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry Ahmed H. El-Khatib Berlin Köster 2014 XI, 122 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: Berlin, Humboldt-Univ., Diss., 2014 Affinitätsmarkierung (DE-588)4293182-4 gnd rswk-swf Proteine (DE-588)4076388-2 gnd rswk-swf Tetraazacyclododecantetraessigsäure 1,4,7,10-Tetraazacyclododecan-1,4,7,10-tetraessigsäure (DE-588)1102901296 gnd rswk-swf Massenspektrometrie (DE-588)4037882-2 gnd rswk-swf Quantitative Analyse (DE-588)4125311-5 gnd rswk-swf Metallkomplexe (DE-588)4038898-0 gnd rswk-swf Peptide (DE-588)4045125-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Peptide (DE-588)4045125-2 s Proteine (DE-588)4076388-2 s Quantitative Analyse (DE-588)4125311-5 s Affinitätsmarkierung (DE-588)4293182-4 s Metallkomplexe (DE-588)4038898-0 s Tetraazacyclododecantetraessigsäure 1,4,7,10-Tetraazacyclododecan-1,4,7,10-tetraessigsäure (DE-588)1102901296 s Massenspektrometrie (DE-588)4037882-2 s DE-604 X:MVB text/html http://deposit.dnb.de/cgi-bin/dokserv?id=4668676&prov=M&dok_var=1&dok_ext=htm Inhaltstext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027379291&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | El-Khatib, Ahmed H. 1979- Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry Affinitätsmarkierung (DE-588)4293182-4 gnd Proteine (DE-588)4076388-2 gnd Tetraazacyclododecantetraessigsäure 1,4,7,10-Tetraazacyclododecan-1,4,7,10-tetraessigsäure (DE-588)1102901296 gnd Massenspektrometrie (DE-588)4037882-2 gnd Quantitative Analyse (DE-588)4125311-5 gnd Metallkomplexe (DE-588)4038898-0 gnd Peptide (DE-588)4045125-2 gnd |
| subject_GND | (DE-588)4293182-4 (DE-588)4076388-2 (DE-588)1102901296 (DE-588)4037882-2 (DE-588)4125311-5 (DE-588)4038898-0 (DE-588)4045125-2 (DE-588)4113937-9 |
| title | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry |
| title_auth | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry |
| title_exact_search | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry |
| title_full | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry Ahmed H. El-Khatib |
| title_fullStr | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry Ahmed H. El-Khatib |
| title_full_unstemmed | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry Ahmed H. El-Khatib |
| title_short | Quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry |
| title_sort | quantitative analysis of chemically modified biopolymers using separation techniques and mass spectrometry |
| topic | Affinitätsmarkierung (DE-588)4293182-4 gnd Proteine (DE-588)4076388-2 gnd Tetraazacyclododecantetraessigsäure 1,4,7,10-Tetraazacyclododecan-1,4,7,10-tetraessigsäure (DE-588)1102901296 gnd Massenspektrometrie (DE-588)4037882-2 gnd Quantitative Analyse (DE-588)4125311-5 gnd Metallkomplexe (DE-588)4038898-0 gnd Peptide (DE-588)4045125-2 gnd |
| topic_facet | Affinitätsmarkierung Proteine Tetraazacyclododecantetraessigsäure 1,4,7,10-Tetraazacyclododecan-1,4,7,10-tetraessigsäure Massenspektrometrie Quantitative Analyse Metallkomplexe Peptide Hochschulschrift |
| url | http://deposit.dnb.de/cgi-bin/dokserv?id=4668676&prov=M&dok_var=1&dok_ext=htm http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027379291&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT elkhatibahmedh quantitativeanalysisofchemicallymodifiedbiopolymersusingseparationtechniquesandmassspectrometry |