The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.
Direct Analysis and Identification of Proteins in Mixtures by LC/MS/MS and Database Searching at the Low-Femtomole Level
Abstract
A method to directly identify proteins contained in mixtures by microcolumn reversed-phase liquid chromatography electrospray ionization tandem mass spectrometry (LC/MS/MS) is studied. In this method, the mixture of proteins is digested with a proteolytic enzyme to produce a large collection of peptides. The complex peptide mixture is then separated on-line with a tandem mass spectrometer, acquiring large numbers of tandem mass spectra. The tandem mass spectra are then used to search a protein database to identify the proteins present. Results from standard protein mixtures show that proteins present in simple mixtures can be readily identified with a 30-fold difference in molar quantity, that the identifications are reproducible, and that proteins within the mixture can be identified at low femtomole levels. Based on these studies, methodology has been developed for direct LC/MS/MS analysis of proteins enriched by immunoaffinity precipitation, specific interaction with a protein-protein fusion product, and specific interaction with a macromolecular complex. The approach described in this article provides a rapid method for the direct identification of proteins in mixtures.
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Bradford, M. M. (1976) · Analytical Biochemistry · doi:10.1016/0003-2697(76)90527-3 · Cited by 255,302
Dynamic instability of microtubule growth
Mitchison, T., & Kirschner, M. (1984) · Nature · doi:10.1038/312237a0 · Cited by 3,337
Cited by
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
Self-citationBest practices and benchmarks for intact protein analysis for top-down mass spectrometry
Donnelly, D. P., Rawlins, C. M., DeHart, C. J., Fornelli, L., Schachner, L. F., Lin, Z., Lippens, J. L., Aluri, K. C., Sarin, R., Chen, B., et al. (2019) · Nature Methods · doi:10.1038/s41592-019-0457-0 · Cited by 377
Yu, Y.-Q., Gilar, M., Lee, P. J., Bouvier, E. S. P., & Gebler, J. C. (2003) · Analytical Chemistry · doi:10.1021/ac0346196 · Cited by 313
Krokhin, O. V., Craig, R., Spicer, V., Ens, W., Standing, K. G., Beavis, R. C., & Wilkins, J. A. (2004) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m400031-mcp200 · Cited by 284
Martin, S. E., Shabanowitz, J., Hunt, D. F., & Marto, J. A. (2000) · Analytical Chemistry · doi:10.1021/ac000497v · Cited by 276
Metadata
- Authors
- McCormack, A. L., Schieltz, D. M., Goode, B., Yang, S., Barnes, G., Drubin, D., & Yates, J. R.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1997-02-01
- Volume
- 69
- Issue
- 4
- Pages
- 767-776
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W1965383843
- Cited by
- 523
- References
- 2
- FWCI
- 14.09
- Type
- journal article
- Language
- en
- Open access
- closed
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- The paper
- McCormack, A. L., Schieltz, D. M., Goode, B., Yang, S., Barnes, G., Drubin, D., & Yates, J. R. (1997). Direct Analysis and Identification of Proteins in Mixtures by LC/MS/MS and Database Searching at the Low-Femtomole Level. Analytical Chemistry. https://doi.org/10.1021/ac960799q
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