Quantitative Proteomics Using Ultralow Flow Capillary Electrophoresis–Mass Spectrometry
Open accessAbstract
In this work, we evaluate the incorporation of an ultralow flow interface for coupling capillary electrophoresis (CE) and mass spectrometry (MS), in combination with reversed-phase high-pressure liquid chromatography (HPLC) fractionation as an alternate workflow for quantitative proteomics. Proteins, extracted from a SILAC (stable isotope labeling by amino acids in cell culture) labeled and an unlabeled yeast strain were mixed and digested enzymatically in solution. The resulting peptides were fractionated using RP-HPLC and analyzed by CE–MS yielding a total of 28 538 quantified peptides that correspond to 3 272 quantified proteins. CE–MS analysis was performed using a neutral capillary coating, providing the highest separation efficiency at ultralow flow conditions (<10 nL/min). Moreover, we were able to demonstrate that CE–MS is a powerful method for the identification of low-abundance modified peptides within the same sample. Without any further enrichment strategies, we succeeded in quantifying 1 371 phosphopeptides present in the CE–MS data set and found 49 phosphopeptides to be differentially regulated in the two yeast strains. Including acetylation, phosphorylation, deamidation, and oxidized forms, a total of 8 106 modified peptides could be identified in addition to 33 854 unique peptide sequences found. The work presented here shows the first quantitative proteomics approach that combines SILAC labeling with CE–MS analysis.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
A SILAC CE-MS approach quantified 1371 phosphopeptides without enrichment.
“Faserl and co-workers reported a SILAC approach that resulted in a quantification of 1371 phosphopeptides by CE-MS, without using any enrichment strategy127.”
From Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research (2019) · cites this paper as [127]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Global analysis of protein expression in yeast
Ghaemmaghami, S., Huh, W.-K., Bower, K., Howson, R. W., Belle, A., Dephoure, N., O'Shea, E. K., & Weissman, J. S. (2003) · Nature · doi:10.1038/nature02046 · Cited by 3,750 · see citing works
An Automated Multidimensional Protein Identification Technology for Shotgun Proteomics
Wolters, D. A., Washburn, M. P., & Yates, J. R. (2001) · Analytical Chemistry · doi:10.1021/ac010617e · Cited by 1,821 · see citing works
Larsen, M. R., Thingholm, T. E., Jensen, O. N., Roepstorff, P., & Jørgensen, T. J. D. (2005) · Molecular & Cellular Proteomics · doi:10.1074/mcp.t500007-mcp200 · Cited by 1,478 · see citing works
Comprehensive mass-spectrometry-based proteome quantification of haploid versus diploid yeast
de Godoy, L. M. F., Olsen, J. V., Cox, J., Nielsen, M. L., Hubner, N. C., Fröhlich, F., Walther, T. C., & Mann, M. (2008) · Nature · doi:10.1038/nature07341 · Cited by 873 · see citing works
On-line mass spectrometric detection for capillary zone electrophoresis
Olivares, J. A., Nhung, N. T., Yonker, C. R., & Smith, R. D. (1987) · Analytical Chemistry · doi:10.1021/ac00135a034 · Cited by 635 · see citing works
Schmidt, A., Karas, M., & Dülcks, T. (2003) · Journal of the American Society for Mass Spectrometry · doi:10.1016/s1044-0305(03)00128-4 · Cited by 535 · see citing works
Improved electrospray ionization interface for capillary zone electrophoresis-mass spectrometry
Smith, R. D., Barinaga, C. J., & Udseth, H. R. (1988) · Analytical Chemistry · doi:10.1021/ac00169a022 · Cited by 534 · see citing works
Gilár, M., Olivova, P., Daly, A. E., & Gebler, J. C. (2005) · Journal of Separation Science · doi:10.1002/jssc.200500116 · Cited by 427 · see citing works
Endosomal transport function in yeast requires a novel AAA‐type ATPase, Vps4p
Babst, M., Sato, T. K., Banta, L. M., & Emr, S. D. (1997) · The EMBO Journal · doi:10.1093/emboj/16.8.1820 · Cited by 424 · see citing works
Moini, M. (2007) · Analytical Chemistry · doi:10.1021/ac0704560 · Cited by 322 · see citing works
Wojcik, R., Dada, O. O., Sadilek, M., & Dovichi, N. J. (2010) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.4672 · Cited by 234 · see citing works
Maxwell, E. J., & Chen, D. D. Y. (2008) · Analytica Chimica Acta · doi:10.1016/j.aca.2008.06.034 · Cited by 197 · see citing works
Evaluation and Optimization of ZIC-HILIC-RP as an Alternative MudPIT Strategy
Boersema, P. J., Divecha, N., Heck, A. J. R., & Mohammed, S. (2007) · Journal of Proteome Research · doi:10.1021/pr060589m · Cited by 190 · see citing works
Liquid junction coupling for capillary zone electrophoresis/ion spray mass spectrometry
Lee, E. D., Mück, W. M., Henion, J. D., & Covey, T. R. (1989) · Journal of Mass Spectrometry · doi:10.1002/bms.1200180932 · Cited by 180 · see citing works
Intact protein analysis in the biopharmaceutical field
Staub, A., Guillarme, D., Schappler, J., Veuthey, J., & Rudaz, S. L. (2011) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2011.01.031 · Cited by 179 · see citing works
Capillary electrophoresis and isoelectric focusing in peptide and protein analysis
Righetti, P. G., Sebastiano, R., & Citterio, A. (2012) · PROTEOMICS · doi:10.1002/pmic.201200378 · Cited by 166 · see citing works
Busnel, J., Schoenmaker, B., Ramautar, R., Carrasco‐Pancorbo, A., Ratnayake, C. K., Feitelson, J. S., Chapman, J. D., Deelder, A. M., & Mayboroda, O. A. (2010) · Analytical Chemistry · doi:10.1021/ac102159d · Cited by 165 · see citing works
Faserl, K., Sarg, B., Kremser, L., & Lindner, H. H. (2011) · Analytical Chemistry · doi:10.1021/ac2010372 · Cited by 153 · see citing works
Self-citationDecoupling CE and ESI for a more robust interface with MS
Maxwell, E. J., Zhong, X., Zhang, H., van Zeijl, N., & Chen, D. D. Y. (2010) · Electrophoresis · doi:10.1002/elps.200900517 · Cited by 150 · see citing works
Haselberg, R., de Jong, G. J., & Somsen, G. W. (2013) · Analytical Chemistry · doi:10.1021/ac303158f · Cited by 149 · see citing works
Wang, Y., Fonslow, B. R., Wong, C. C. L., Nakorchevsky, A., & Yates, J. R. (2012) · Analytical Chemistry · doi:10.1021/ac301091m · Cited by 144 · see citing works
Li, Y., Champion, M. M., Sun, L., Champion, P. A. D., Wojcik, R., & Dovichi, N. J. (2012) · Analytical Chemistry · doi:10.1021/ac202899p · Cited by 128 · see citing works
Nilsson, S., Wetterhall, M., Bergquist, J., Nyholm, L., & Markides, K. E. (2001) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.466 · Cited by 110 · see citing works
Haselberg, R., Ratnayake, C. K., de Jong, G. J., & Somsen, G. W. (2010) · Journal of Chromatography A · doi:10.1016/j.chroma.2010.10.006 · Cited by 109 · see citing works
Zhu, G., Sun, L., Yan, X., & Dovichi, N. J. (2013) · Analytical Chemistry · doi:10.1021/ac303750g · Cited by 103 · see citing works
Heemskerk, A. A. M., Busnel, J., Schoenmaker, B., Derks, R. J. E., Klychnikov, O. I., Hensbergen, P. J., Deelder, A. M., & Mayboroda, O. A. (2012) · Analytical Chemistry · doi:10.1021/ac300641x · Cited by 100 · see citing works
Sun, L., Hebert, A. S., Yan, X., Zhao, Y., Westphall, M. S., Rush, M. J. P., Zhu, G., Champion, M. M., Coon, J. J., & Dovic̀hi, N. J. (2014) · Angewandte Chemie International Edition · doi:10.1002/anie.201409075 · Cited by 91 · see citing works
Gahoual, R., Burr, A., Busnel, J.-M., Kuhn, L., Hammann, P., Beck, A., François, Y.-N., & Leize-Wagner, E. (2013) · mAbs · doi:10.4161/mabs.23995 · Cited by 90 · see citing works
CE–MS for proteomics: Advances in interface development and application
Ramautar, R., Heemskerk, A. A. M., Hensbergen, P. J., Deelder, A. M., Busnel, J.-M., & Mayboroda, O. A. (2012) · Journal of Proteomics · doi:10.1016/j.jprot.2012.04.050 · Cited by 86 · see citing works
Long-lived metallized tips for nanoliter electrospray mass spectrometry
Valaskovic, G. A., & McLafferty, F. W. (1996) · Journal of the American Society for Mass Spectrometry · doi:10.1016/s1044-0305(96)00152-3 · Cited by 83 · see citing works
Sarg, B., Faserl, K., Kremser, L., Halfinger, B., Sebastiano, R., & Lindner, H. H. (2013) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m112.024109 · Cited by 76 · see citing works
Self-citationLindner, H. H., Helliger, W., Dirschlmayer, A., Jaquemar, M., & Puschendorf, B. (1992) · Biochemical Journal · doi:10.1042/bj2830467 · Cited by 67 · see citing works
Self-citationZhong, X., Maxwell, E. J., Ratnayake, C. K., Mack, S., & Chen, D. D. Y. (2011) · Analytical Chemistry · doi:10.1021/ac202130f · Cited by 66 · see citing works
The Microheterogeneity of the Mammalian H10Histone
Lindner, H. H., Sarg, B., Hoertnagl, B., & Helliger, W. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.21.13324 · Cited by 63 · see citing works
Self-citationRitorto, M. S., Cook, K., Tyagi, K., Pedrioli, P. G. A., & Trost, M. (2013) · Journal of Proteome Research · doi:10.1021/pr301011r · Cited by 61 · see citing works
Sun, L., Zhu, G., & Dovic̀hi, N. J. (2013) · Analytical Chemistry · doi:10.1021/ac400523x · Cited by 55 · see citing works
Gahoual, R., Busnel, J., Wolff, P., François, Y., & Leize‐Wagner, E. (2013) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-013-7226-1 · Cited by 48 · see citing works
Yang, H., Fung, Y. M. E., Zubarev, A. R., et al. (2009) · Journal of Proteome Research · doi:10.1021/pr900428m · Cited by 43 · see citing works
Separation of phosphorylated histone H1 variants by high-performance capillary electrophoresis
Lindner, H. H., Helliger, W., Dirschlmayer, A., Talasz, H., Wurm, M., Sarg, B., Jaquemar, M., & Puschendorf, B. (1992) · Journal of Chromatography A · doi:10.1016/0021-9673(92)87126-s · Cited by 38 · see citing works
Self-citationApplication of high‐performance capillary electrophoresis to the analysis of H1 histones
Lindner, H. H., Wurm, M., Dirschlmayer, A., Sarg, B., & Helliger, W. (1993) · Electrophoresis · doi:10.1002/elps.1150140174 · Cited by 36 · see citing works
Self-citationTestis-specific Linker Histone H1t Is Multiply Phosphorylated during Spermatogenesis
Sarg, B., Chwatal, S., Talasz, H., & Lindner, H. H. (2008) · Journal of Biological Chemistry · doi:10.1074/jbc.m805925200 · Cited by 35 · see citing works
Self-citationAnalysis of histones, histone variants, and their post‐translationally modified forms
Lindner, H. H. (2008) · Electrophoresis · doi:10.1002/elps.200800094 · Cited by 33 · see citing works
Self-citationDifferentiating N-Terminal Aspartic and Isoaspartic Acid Residues in Peptides
Sargaeva, N. P., Lin, C. T., & O’Connor, P. B. (2011) · Analytical Chemistry · doi:10.1021/ac201223d · Cited by 30 · see citing works
Girolamo, F. D., Righetti, P. G., Soste, M., et al. (2013) · Journal of Proteomics · doi:10.1016/j.jprot.2013.05.037 · Cited by 15 · see citing works
Boschetti, E., & Giorgio, P. (2014) · Cited by 13 · see citing works
Lindner, H. H., Sarg, B., & Helliger, W. (2004) · PubMed · Cited by 9 · see citing works
Self-citation
Flagged first, then by citations
Research works printed but not linked: 4
- Lindner H.; Sarg B.; Helliger W. J. Capillary Electrophor. Microchip Technol. 2003, 8, 59– 67. 14596337
- Smith R. D.; Barinaga C. J.; Udseth H. R. Anal. Chem. 1988, 60, 1948– 1952.
- Lee E. D.; Muck W.; Henion J. D.; Covey T. R. Biomed. Environ. Mass Spectrom. 1989, 18, 844– 850.
- Olivares J. A.; Nguyen N. T.; Yonker C. R.; Smith R. D. Anal. Chem. 1987, 59, 1230– 1232.
Cited by
Sorted by citations
Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research
Gomes, F. P., & Yates, J. R. (2019) · Mass Spectrometry Reviews · doi:10.1002/mas.21599 · Cited by 112 · see citing works
Lubeckyj, R. A., McCool, E. N., Shen, X., Kou, Q., Liu, X., & Sun, L. (2017) · Analytical Chemistry · doi:10.1021/acs.analchem.7b02532 · Cited by 96 · see citing works
Capillary zone electrophoresis-mass spectrometry for bottom-up proteomics
Zhang, Z., Qu, Y., & Dovichi, N. J. (2018) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2018.08.008 · Cited by 61 · see citing works
Ludwig, K. R., Sun, L., Zhu, G., Dovichi, N. J., & Hummon, A. B. (2015) · Analytical Chemistry · doi:10.1021/acs.analchem.5b02457 · Cited by 56 · see citing works
Predicting Electrophoretic Mobility of Tryptic Peptides for High-Throughput CZE-MS Analysis
Krokhin, O. V., Anderson, G., Spicer, V., Sun, L., & Dovichi, N. J. (2017) · Analytical Chemistry · doi:10.1021/acs.analchem.6b04544 · Cited by 53 · see citing works
Guo, X., Fillmore, T. L., Gao, Y., & Tang, K. (2016) · Analytical Chemistry · doi:10.1021/acs.analchem.5b04912 · Cited by 54 · see citing works
Zhu, G., Sun, L., & Dovichi, N. J. (2016) · The Analyst · doi:10.1039/c6an01140c · Cited by 52 · see citing works
Chen, D., Shen, X., & Sun, L. (2018) · Analytica Chimica Acta · doi:10.1016/j.aca.2018.01.037 · Cited by 47 · see citing works
- Authors
- Faserl, K., Kremser, L., Müller, M., Teis, D., & Lindner, H. H.
- Affiliations
- Innsbruck Medical University (AT)
- Innsbruck Medical University , , Innrain 80-82 , ,
- Corresponding author
- Lindner, Herbert H.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2015-04-03
- Volume
- 87
- Issue
- 9
- Pages
- 4633-4640
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- PMC
- PMC4423236
- OpenAlex
- W1985017503
- Cited by
- 57
- References
- 46
- Citations by year
- 2026: 2 · 2024: 1 · 2023: 2 · 2022: 3 · 2021: 2 · 2020: 2 · 2019: 7 · 2018: 13 · 2017: 11 · 2016: 9 · 2015: 4 · 2013: 1
- FWCI
- 5.49
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Analytical Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Austrian Science Fund (Y444-B12)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Keywords
- quantitative proteomics, capillary electrophoresis-mass spectrometry, yeast proteome, phosphopeptides, modified peptides, peptide fractionation, reversed-phase HPLC, protein quantification, differential regulation
- MeSH terms
- Chromatography, Liquid, Peptides, Proteins, Mass Spectrometry, Electrophoresis, Capillary, Proteome, Proteomics
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review