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Enhanced detection and identification of multiply phosphorylated peptides using TiO₂ enrichment in combination with MALDI TOF/TOF MS
The analysis of PTMs such as phosphorylation has become an important field in MS because they can directly indicate protein states and interactions. Whereas the characterization of singly and doubly phosphorylated peptides has almost become routine, identifying phosphorylation events at multiple res...
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Published in: | Proteomics (Weinheim) 2008-11, Vol.8 (21), p.4577-4592 |
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container_title | Proteomics (Weinheim) |
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creator | Schmidt, Andreas Csaszar, Edina Ammerer, Gustav Mechtler, Karl |
description | The analysis of PTMs such as phosphorylation has become an important field in MS because they can directly indicate protein states and interactions. Whereas the characterization of singly and doubly phosphorylated peptides has almost become routine, identifying phosphorylation events at multiple residues within a small region of a protein is still problematic. The identification of multiple modifications can be further hampered by low sequence information due to multiple neutral losses from phosphorylated side chains. Here we present a strategy for the analysis of complex phosphopeptides that combines peptide enrichment by titanium dioxide, separation by RP separation on monolithic columns and MS using high energy HE-CAD in a MALDI TOF/TOF analyser. Using synthetic phosphopeptides our approach is compared to multistage activation (MSA) MS/MS and the recently described electron transfer dissociation (ETD) method using an ESI-LTQ mass spectrometer. |
doi_str_mv | 10.1002/pmic.200800279 |
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Whereas the characterization of singly and doubly phosphorylated peptides has almost become routine, identifying phosphorylation events at multiple residues within a small region of a protein is still problematic. The identification of multiple modifications can be further hampered by low sequence information due to multiple neutral losses from phosphorylated side chains. Here we present a strategy for the analysis of complex phosphopeptides that combines peptide enrichment by titanium dioxide, separation by RP separation on monolithic columns and MS using high energy HE-CAD in a MALDI TOF/TOF analyser. Using synthetic phosphopeptides our approach is compared to multistage activation (MSA) MS/MS and the recently described electron transfer dissociation (ETD) method using an ESI-LTQ mass spectrometer.</description><identifier>ISSN: 1615-9853</identifier><identifier>EISSN: 1615-9861</identifier><identifier>DOI: 10.1002/pmic.200800279</identifier><identifier>PMID: 18972529</identifier><language>eng</language><publisher>Weinheim: Wiley-VCH Verlag</publisher><subject>Amino Acid Sequence ; Chromatographic enrichment ; Chromatography, High Pressure Liquid ; Crystallization ; Indicators and Reagents ; Isotope Labeling ; Mass spectrometry ; Molecular Sequence Data ; Osteopontin - analysis ; Phosphopeptides ; Phosphopeptides - analysis ; Phosphoproteomics ; Phosphorylation ; Proteomics ; Spectrometry, Mass, Electrospray Ionization ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tandem Mass Spectrometry ; Titanium</subject><ispartof>Proteomics (Weinheim), 2008-11, Vol.8 (21), p.4577-4592</ispartof><rights>Copyright © 2008 WILEY‐VCH Verlag GmbH & Co. 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Using synthetic phosphopeptides our approach is compared to multistage activation (MSA) MS/MS and the recently described electron transfer dissociation (ETD) method using an ESI-LTQ mass spectrometer.</description><subject>Amino Acid Sequence</subject><subject>Chromatographic enrichment</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Crystallization</subject><subject>Indicators and Reagents</subject><subject>Isotope Labeling</subject><subject>Mass spectrometry</subject><subject>Molecular Sequence Data</subject><subject>Osteopontin - analysis</subject><subject>Phosphopeptides</subject><subject>Phosphopeptides - analysis</subject><subject>Phosphoproteomics</subject><subject>Phosphorylation</subject><subject>Proteomics</subject><subject>Spectrometry, Mass, Electrospray Ionization</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><subject>Tandem Mass Spectrometry</subject><subject>Titanium</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><recordid>eNpFkTtPIzEUhS20iHdLueuKLuDHOLZLFF6REgWJUFuO55oYzXhmxzNCaSn4oftL1hA2W1xdX_s7R1c-CJ1TckkJYVdtHdwlI0TlQeo9dETHVIy0GtMfu7Pgh-g4pVdCqFRaHqBDmhsTTB-hj9u4ttFBiUvowfWhidjGEocSYh98cPbrqvG4Hqo-tNUGt-sm5eo2le2zroW2z3TCQwrxBS_D4s_7O4bYBbeuswkOEbumXoW4tXoL_RrPr2c3U7xc3F3lwvOnU7TvbZXg7LufoOe72-XkYTRb3E8n17ORZ4rrkSicA2q1pso5WxacSFUUxPGSeADtpGdaMMktpVCsJPeEeqmYBcZBKGf5CbrY-rZd83uA1Js6JAdVZSM0QzJjLQuhBc_gz29wWNVQmrYLte025t_PZUBvgbdQweb_OzGfuZjPXMwuF_M4n052U9b-2mq9bYx96UIyz0-MUE6oECovwP8CzkiM9w</recordid><startdate>20081101</startdate><enddate>20081101</enddate><creator>Schmidt, Andreas</creator><creator>Csaszar, Edina</creator><creator>Ammerer, Gustav</creator><creator>Mechtler, Karl</creator><general>Wiley-VCH Verlag</general><general>WILEY‐VCH Verlag</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20081101</creationdate><title>Enhanced detection and identification of multiply phosphorylated peptides using TiO₂ enrichment in combination with MALDI TOF/TOF MS</title><author>Schmidt, Andreas ; Csaszar, Edina ; Ammerer, Gustav ; Mechtler, Karl</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f2839-54cce1a9918ccad43078440c3d0fee9c7f295273a11e4b73f01f782ae23e58ca3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Amino Acid Sequence</topic><topic>Chromatographic enrichment</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Crystallization</topic><topic>Indicators and Reagents</topic><topic>Isotope Labeling</topic><topic>Mass spectrometry</topic><topic>Molecular Sequence Data</topic><topic>Osteopontin - analysis</topic><topic>Phosphopeptides</topic><topic>Phosphopeptides - analysis</topic><topic>Phosphoproteomics</topic><topic>Phosphorylation</topic><topic>Proteomics</topic><topic>Spectrometry, Mass, Electrospray Ionization</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>Tandem Mass Spectrometry</topic><topic>Titanium</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schmidt, Andreas</creatorcontrib><creatorcontrib>Csaszar, Edina</creatorcontrib><creatorcontrib>Ammerer, Gustav</creatorcontrib><creatorcontrib>Mechtler, Karl</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schmidt, Andreas</au><au>Csaszar, Edina</au><au>Ammerer, Gustav</au><au>Mechtler, Karl</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enhanced detection and identification of multiply phosphorylated peptides using TiO₂ enrichment in combination with MALDI TOF/TOF MS</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2008-11-01</date><risdate>2008</risdate><volume>8</volume><issue>21</issue><spage>4577</spage><epage>4592</epage><pages>4577-4592</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>The analysis of PTMs such as phosphorylation has become an important field in MS because they can directly indicate protein states and interactions. Whereas the characterization of singly and doubly phosphorylated peptides has almost become routine, identifying phosphorylation events at multiple residues within a small region of a protein is still problematic. The identification of multiple modifications can be further hampered by low sequence information due to multiple neutral losses from phosphorylated side chains. Here we present a strategy for the analysis of complex phosphopeptides that combines peptide enrichment by titanium dioxide, separation by RP separation on monolithic columns and MS using high energy HE-CAD in a MALDI TOF/TOF analyser. Using synthetic phosphopeptides our approach is compared to multistage activation (MSA) MS/MS and the recently described electron transfer dissociation (ETD) method using an ESI-LTQ mass spectrometer.</abstract><cop>Weinheim</cop><pub>Wiley-VCH Verlag</pub><pmid>18972529</pmid><doi>10.1002/pmic.200800279</doi><tpages>16</tpages></addata></record> |
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subjects | Amino Acid Sequence Chromatographic enrichment Chromatography, High Pressure Liquid Crystallization Indicators and Reagents Isotope Labeling Mass spectrometry Molecular Sequence Data Osteopontin - analysis Phosphopeptides Phosphopeptides - analysis Phosphoproteomics Phosphorylation Proteomics Spectrometry, Mass, Electrospray Ionization Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Tandem Mass Spectrometry Titanium |
title | Enhanced detection and identification of multiply phosphorylated peptides using TiO₂ enrichment in combination with MALDI TOF/TOF MS |
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