| Nucleic acids in protein samples interfere with phosphopeptide identification by immobilized-metal-ion affinity chromatography and mass spectrometry. | |
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MedLine Citation:
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PMID: 19412670 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Immobilized-metal-ion affinity chromatography (IMAC) is used extensively for phosphopeptide enrichment in phosphoproteomics. However, the effect of nucleic acids in protein samples on phosphopeptide enrichment by IMAC has not yet been well clarified. In this study, we demonstrate that IMAC beads possess a strong adsorption of nucleic acids, especially single-stranded or single-stranded-region-containing nucleic acids, leading to approximately 50% loss of phosphopeptides during the process of IMAC enrichment. Therefore, nucleic acids must be removed from protein samples prior to IMAC. Acetonitrile (ACN) precipitation, a simple and efficient procedure, was established to remove nucleic acids from the protein samples. We showed that ACN precipitation approximately doubled the phosphopeptide number identified by IMAC and mass spectrometry, indicating that nucleic acid removal significantly improves the identification of phosphopeptides. |
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Authors:
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Yaojun Li; Yuanming Luo; Shuzhen Wu; Youhe Gao; Yanxin Liu; Dexian Zheng |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-05-02 |
Journal Detail:
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Title: Molecular biotechnology Volume: 43 ISSN: 1073-6085 ISO Abbreviation: Mol. Biotechnol. Publication Date: 2009 Sep |
Date Detail:
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Created Date: 2009-08-21 Completed Date: 2009-10-22 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9423533 Medline TA: Mol Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 59-66 Citation Subset: IM |
Affiliation:
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National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, 5 Dong Dan San Tiao, Beijing 100005, China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adsorption Artifacts Chromatography, Affinity / methods* Complex Mixtures / analysis, chemistry Mass Spectrometry / methods* Metals / chemistry Nucleic Acids / analysis*, chemistry* Phosphopeptides / analysis*, chemistry* Proteins / chemistry* |
| Chemical | |
Reg. No./Substance:
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0/Complex Mixtures; 0/Metals; 0/Nucleic Acids; 0/Phosphopeptides; 0/Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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