Proteomic profiling of intact proteins using WAX-RPLC 2-D separations and FTICR mass spectrometry
We investigated the combination of weak anion exchange (WAX) fractionation and on-line reversed-phase liquid chromatography (RPLC) separation using a 12 T FTICR mass spectrometer for the detection of intact proteins from a Shewanella oneidensis MR-1 cell lysate. This work aimed at optimizing intact protein detection for profiling proteins at a level that incorporates their modification state. A total of 715 intact proteins were detected, and the combined results from the WAX fractions and the unfractionated cell lysate were aligned using LC-MS features to facilitate protein abundance measurements. Protein identifications and post-translational modifications were assigned for approximately 10% of the detected proteins by comparing intact protein mass measurements to proteins identified in peptide MS/MS analysis of an aliquot of the same fraction. Intact proteins were also detected for S. oneidensis lysates obtained from cells grown on 13C-, 15N-depleted media under aerobic and sub-oxic conditions. The strategy can be readily applied for measuring differential protein abundances and provides a platform for high-throughput selection of biologically relevant targets for further characterization.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2007 |
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| Erschienen: |
2007 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:6 |
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| Enthalten in: |
Journal of proteome research - 6(2007), 2 vom: 27. Feb., Seite 602-10 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Sharma, Seema [VerfasserIn] |
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| Themen: |
Bacterial Proteins |
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| Anmerkungen: |
Date Completed 03.04.2007 Date Revised 03.12.2007 published: Print Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM168122898 |
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| 100 | 1 | |a Sharma, Seema |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Proteomic profiling of intact proteins using WAX-RPLC 2-D separations and FTICR mass spectrometry |
| 264 | 1 | |c 2007 | |
| 336 | |a Text |b txt |2 rdacontent | ||
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| 500 | |a Date Completed 03.04.2007 | ||
| 500 | |a Date Revised 03.12.2007 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a We investigated the combination of weak anion exchange (WAX) fractionation and on-line reversed-phase liquid chromatography (RPLC) separation using a 12 T FTICR mass spectrometer for the detection of intact proteins from a Shewanella oneidensis MR-1 cell lysate. This work aimed at optimizing intact protein detection for profiling proteins at a level that incorporates their modification state. A total of 715 intact proteins were detected, and the combined results from the WAX fractions and the unfractionated cell lysate were aligned using LC-MS features to facilitate protein abundance measurements. Protein identifications and post-translational modifications were assigned for approximately 10% of the detected proteins by comparing intact protein mass measurements to proteins identified in peptide MS/MS analysis of an aliquot of the same fraction. Intact proteins were also detected for S. oneidensis lysates obtained from cells grown on 13C-, 15N-depleted media under aerobic and sub-oxic conditions. The strategy can be readily applied for measuring differential protein abundances and provides a platform for high-throughput selection of biologically relevant targets for further characterization | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 7 | |a Bacterial Proteins |2 NLM | |
| 700 | 1 | |a Simpson, David C |e verfasserin |4 aut | |
| 700 | 1 | |a Tolić, Nikola |e verfasserin |4 aut | |
| 700 | 1 | |a Jaitly, Navdeep |e verfasserin |4 aut | |
| 700 | 1 | |a Mayampurath, Anoop M |e verfasserin |4 aut | |
| 700 | 1 | |a Smith, Richard D |e verfasserin |4 aut | |
| 700 | 1 | |a Pasa-Tolić, Ljiljana |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Journal of proteome research |d 2002 |g 6(2007), 2 vom: 27. Feb., Seite 602-10 |w (DE-627)NLM124173470 |x 1535-3907 |7 nnns |
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