The DOI matches its Crossref record; 12 of the 12 references it cites were found in a registry or an index.
Selective Isolation at the Femtomole Level of Phosphopeptides from Proteolytic Digests Using 2D-NanoLC-ESI-MS/MS and Titanium Oxide Precolumns
Abstract
Selective detection of phosphopeptides from proteolytic digests is a challenging and highly relevant task in many proteomics applications. Often phosphopeptides are present in small amounts and need selective isolation or enrichment before identification. Here we report a novel automated method for the enrichment of phosphopeptides from complex mixtures. The method employs a two-dimensional column setup, with titanium oxide-based solid-phase material (Titansphere) as the first dimension and reversed-phase material as the second dimension. Phosphopeptides are separated from nonphosphorylated peptides by trapping them under acidic conditions on a TiO(2) precolumn. Nonphosphorylated peptides break through and are trapped on a reversed-phase precolumn after which they are analyzed by nanoflow LC-ESI-MS/MS. Subsequently, phosphopeptides are desorbed from the TiO(2) column under alkaline conditions, reconcentrated onto the reversed-phase precolumn, and analyzed by nanoflow LC-ESI-MS/MS. The selectivity and practicality of using TiO(2) precolumns for trapping phosphopeptides are demonstrated via the analysis of a model peptide RKISASEF, in a 1:1 mixture of a non- and a monophosphorylated form. A sample of 125 fmol of the phosphorylated peptide could easily be isolated from the nonphosphorylated peptide with a recovery above 90%. In addition, proteolytic digests of three different autophosphorylation forms of the 153-kDa homodimeric cGMP-dependent protein kinase are analyzed. From proteolytic digests of the fully autophosphorylated protein at least eight phosphorylation sites are identified, including two previously uncharacterized sites, namely, Ser-26 and Ser-44. Ser-26 is characterized as a minor phosphorylation site in purified PKG samples, while Ser-44 is identified as a novel in vitro autophosphorylation target. These results clearly show that TiO(2) has strong affinity for phosphorylated peptides, and thus, we conclude that this material has a high potential in the field of phosphoproteomics.
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Cites
Hunter, T. (2000) · Cell · doi:10.1016/s0092-8674(00)81688-8 · Cited by 2,586
Phosphoproteome analysis by mass spectrometry and its application to Saccharomyces cerevisiae
Ficarro, S. B., McCleland, M. L., Stukenberg, P. T., Burke, D. J., Ross, M. M., Shabanowitz, J., Hunt, D. F., & White, F. M. (2002) · Nature Biotechnology · doi:10.1038/nbt0302-301 · Cited by 1,636
Immobilized Gallium(III) Affinity Chromatography of Phosphopeptides
Posewitz, M. C., & Tempst, P. (1999) · Analytical Chemistry · doi:10.1021/ac981409y · Cited by 832
Nühse, T. S., Stensballe, A., Jensen, O. N., & Peck, S. C. (2003) · Molecular & Cellular Proteomics · doi:10.1074/mcp.t300006-mcp200 · Cited by 545
Pandey, A., Podtelejnikov, A. V., Blagoev, B., Bustelo, X. R., Mann, M., & Lodish, H. F. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.97.1.179 · Cited by 406
Analysis of phosphorylated proteins and peptides by mass spectrometry
McLachlin, D. T., & Chait, B. T. (2001) · Current Opinion in Chemical Biology · doi:10.1016/s1367-5931(00)00250-7 · Cited by 382
Nanoscale LC-MS(n): technical design and applications to peptide and protein analysis
Meiring, H. D., van der Heeft, E., ten Hove, G. J., & de Jong, A. P. J. M. (2002) · Journal of Separation Science · doi:10.1002/1615-9314(20020601)25:9<557::aid-jssc557>3.0.co;2-f · Cited by 267
Huddleston, M. J., Annan, R. S., Bean, M. F., & Carr, S. A. (1993) · Journal of the American Society for Mass Spectrometry · doi:10.1016/1044-0305(93)80049-5 · Cited by 242
Haydon, C. E., Eyers, P. A., Aveline-Wolf, L. D., Resing, K. A., Maller, J. L., & Ahn, N. G. (2003) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m300054-mcp200 · Cited by 137
Selective Enrichment of Phospholipids by Titania
Ikeguchi, Y., & Nakamura, H. (2000) · Analytical Sciences · doi:10.2116/analsci.16.541 · Cited by 78
van der Heeft, E., ten Hove, G. J., Herberts, C. A., Meiring, H. D., van Els, C. A. C. M., & de Jong, A. P. J. M. (1998) · Analytical Chemistry · doi:10.1021/ac9801014 · Cited by 64
Cloning cell cycle regulatory genes by transcomplementation in yeast
Norbury, C. J., & Moreno, S. (1997) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(97)83006-6 · Cited by 41
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Degterev, A., Hitomi, J., Germscheid, M., Ch'en, I. L., Korkina, O., Teng, X., Abbott, D., Cuny, G. D., Yuan, C., Wagner, G., et al. (2008) · Nature Chemical Biology · doi:10.1038/nchembio.83 · Cited by 2,059
Reddy, L. H., Arias, J. L., Nicolas, J., & Couvreur, P. (2012) · Chemical Reviews · doi:10.1021/cr300068p · Cited by 2,010
A tissue-specific atlas of mouse protein phosphorylation and expression
Huttlin, E. L., Jedrychowski, M. P., Elias, J. E., Goswami, T., Rad, R., Beausoleil, S. A., Villén, J., Haas, W., Sowa, M. E., & Gygi, S. P. (2010) · Cell · doi:10.1016/j.cell.2010.12.001 · Cited by 1,778
A quantitative atlas of mitotic phosphorylation
Dephoure, N., Zhou, C., Villén, J., Beausoleil, S. A., Bakalarski, C. E., Elledge, S. J., & Gygi, S. P. (2008) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0805139105 · Cited by 1,608
Protein Analysis by Shotgun/Bottom-up Proteomics
Zhang, Y., Fonslow, B. R., Shan, B., Baek, M.-C., & Yates, J. R. (2013) · Chemical Reviews · doi:10.1021/cr3003533 · Cited by 1,491
Metadata
- Authors
- Pinkse, M. W. H., Uitto, P. M., Hilhorst, M. J., Ooms, B., & Heck, A. J. R.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2004-06-15
- Volume
- 76
- Issue
- 14
- Pages
- 3935-3943
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2033889320
- Cited by
- 891
- References
- 12
- FWCI
- 18.39
- Type
- journal article
- Language
- en
- Open access
- closed
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