Structural characterization of protein tryptic peptides via liquid chromatography/mass spectrometry and collision-induced dissociation of their doubly charged molecular ions
Abstract
The formation of multiply charged molecular ions via the field-assisted ion evaporation mechanism during electrospray ionization enables the use of an atmospheric pressure ionization quadrupole mass spectrometer system for characterizing biologically important peptides. The straightforward implementation of high-performance liquid chromatography (HPLC) into this new strategy to determine the molecular weight of tryptic peptides via the pneumatically assisted electrospray (ion spray) interface is presented. Examples utilizing both microbore (1.0 mm) and standard bore (4.6 mm) inside diameter columns are shown for the LC/MS molecular weight determination of tryptic peptides in methionyl-human growth hormone (met-hGH). Injected levels from 50 to 75 pmol of tryptic digest onto 1 mm i.d. HPLC columns provided full-scan LC/MS or LC/MS/MS results without postcolumn splitting of the effluent. When standard 4.6 mm i.d. HPLC columns were used, a 20:1 postcolumn split was utilized, which required from 1 to 5 nmol of injected tryptic digest for full-scan LC/MS or LC/MS/MS results. Collision-induced dissociation (CID) mass spectra resulting from either "infusion" or on-line LC/MS/MS analysis of the abundant doubly charged ions that predominate for tryptic peptides under electrospray conditions provided structurally useful sequence information for met-hGH and human hemoglobin tryptic digests. The slower mass spectrometer scan rate used during infusion of sample provides more accurate mass assignments than on-line LC/MS or LC/MS/MS, but the latter on-line experiments preclude ambiguities caused by matrix or component interferences. However, in some instances very weak CID product ions preclude complete tryptic peptide structural characterization based upon the CID data alone.(ABSTRACT TRUNCATED AT 250 WORDS)
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
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?
The use of on-line liquid chromatography (LC)-MS makes it possible to determine the molecular weight of each peptide, even when HPLC column fractions contain more than one peptide.
“The use of on-line liquid chromatography (LC)-MS makes it possible to determine the molecular weight of each peptide, even when HPLC column fractions contain more than one peptide (7, 36).”
From The extracellular PI-type proteinase of Lactococcus lactis hydrolyzes beta-casein into more than one hundred different oligopeptides (1995) · cites this paper as [7]
Cites
Electrospray Ionization for Mass Spectrometry of Large Biomolecules
Fenn, J. B., Mann, M., Meng, C. K., Wong, S. F., & Whitehouse, C. M. (1989) · Science · doi:10.1126/science.2675315 · Cited by 7,634 · see citing works
Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons.
Karas, M., & Hillenkamp, F. (1988) · Analytical Chemistry · doi:10.1021/ac00171a028 · Cited by 5,145 · see citing works
Protein and polymer analyses up to m/z 100 000 by laser ionization time-of-flight mass spectrometry
Tanaka, K., Waki, H., Ido, Y., Akita, S., Yoshida, Y., Yoshida, T., & Matsuo, T. (1988) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290020802 · Cited by 3,005 · see citing works
Electrospray interface for liquid chromatographs and mass spectrometers.
Whitehouse, C., Dreyer, R., Yamashita, M., & Fenn, J. (1985) · Analytical Chemistry · doi:10.1021/ac00280a023 · Cited by 1,368 · see citing works
On the evaporation of small ions from charged droplets
Iribarne, J. V., & Thomson, B. A. (1976) · The Journal of Chemical Physics · doi:10.1063/1.432536 · Cited by 1,287 · see citing works
Protein sequencing by tandem mass spectrometry.
Hunt, D. F., Yates, J. R., Shabanowitz, J., Winston, S., & Hauer, C. (1986) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.83.17.6233 · Cited by 1,189 · see citing works
New developments in biochemical mass spectrometry: electrospray ionization.
Smith, R. D., Loo, J., Edmonds, C. G., Barinaga, C., & Udseth, H. (1990) · Analytical Chemistry · doi:10.1021/ac00208a002 · Cited by 975 · see citing works
Bruins, A. P., Covey, T. R., & Henion, J. D. (1987) · Analytical Chemistry · doi:10.1021/ac00149a003 · Cited by 880 · see citing works
Self-citationField induced ion evaporation from liquid surfaces at atmospheric pressure
Thomson, B. A., & Iribarne, J. V. (1979) · The Journal of Chemical Physics · doi:10.1063/1.438198 · Cited by 635 · see citing works
Covey, T. R., Bonner, R. F., Shushan, B. I., Henion, J. D., & Boyd, R. K. (1988) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290021111 · Cited by 593 · see citing works
Self-citationInterpreting mass spectra of multiply charged ions
Mann, M., Meng, C. K., & Fenn, J. B. (1989) · Analytical Chemistry · doi:10.1021/ac00190a023 · Cited by 586 · see citing works
Mass spectrometric determination of the amino acid sequence of peptides and proteins
BIEMANN, K., & Martin, S. A. (1987) · Mass Spectrometry Reviews · doi:10.1002/mas.1280060102 · Cited by 554 · see citing works
Liquid chromatography/mass spectrometry
Covey, T. R., Lee, E. D., Bruins, A. P., & Henion, J. D. (1986) · Analytical Chemistry · doi:10.1021/ac00127a001 · Cited by 472 · see citing works
Self-citationMultiple charging in electrospray ionization of poly(ethylene glycols)
Wong, S. F., Meng, C. K., & Fenn, J. B. (1988) · The Journal of Physical Chemistry · doi:10.1021/j100313a058 · Cited by 392 · see citing works
Loo, J. A., Udseth, H. R., & Smith, R. (1989) · Analytical Biochemistry · doi:10.1016/0003-2697(89)90153-x · Cited by 264 · see citing works
Gas-phase ion decomposition occurring remote to a charge site
Jensen, N. J., Tomer, K. B., & Gross, M. L. (1985) · Journal of the American Chemical Society · doi:10.1021/ja00293a010 · Cited by 260 · see citing works
Meng, C. K., Mann, M., & Fenn, J. B. (1988) · Zeitschrift für Physik D Atoms Molecules and Clusters · doi:10.1007/bf01384871 · Cited by 257 · see citing works
Smith, R., Loa, J. A., Barinaga, C. J., et al. (1990) · Journal of the American Society for Mass Spectrometry · doi:10.1016/1044-0305(90)80006-9 · Cited by 244 · see citing works
Loo, J. A., Udseth, H. R., Smith, R. D., & Futrell, J. H. (1988) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290021006 · Cited by 241 · see citing works
Lee, E. D., Mück, W. M., Henion, J. D., & Covey, T. R. (1988) · Journal of Chromatography A · doi:10.1016/s0021-9673(00)90575-2 · Cited by 209 · see citing works
Self-citationFourier-transform mass spectrometry of large molecules by electrospray ionization.
Henry, K. D., Williams, E. R., Wang, B. H., McLafferty, F., Shabanowitz, J., & Hunt, D. F. (1989) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.86.23.9075 · Cited by 193 · see citing works
High-Performance Liquid Chromatography-Mass Spectrometry
Vestal, M. L. (1984) · Science · doi:10.1126/science.6385251 · Cited by 180 · see citing works
Johnson, R. S., & BIEMANN, K. (1987) · Biochemistry · doi:10.1021/bi00379a001 · Cited by 167 · see citing works
Thomson, B. A., Iribarne, J. V., & Dziedzic, P. J. (1982) · Analytical Chemistry · doi:10.1021/ac00250a020 · Cited by 146 · see citing works
Bruins, A. P., Weidolf, L. O. G., Henion, J. D., et al. (1987) · Analytical Chemistry · doi:10.1021/ac00149a004 · Cited by 133 · see citing works
Self-citationIon‐spray mass spectrometry of marine neurotoxins
Quilliam, M. A., Thomson, B. A., Scott, G. J., et al. (1989) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290030508 · Cited by 130 · see citing works
Barinaga, C. J., Edmonds, C. G., Udseth, H. R., et al. (1989) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290030512 · Cited by 117 · see citing works
Molecular Weight Determinations of Proteins by Californium Plasma Desorption Mass Spectrometry
Sundqvist, B., Roepstorff, P., Fohlman, J., et al. (1984) · Science · doi:10.1126/science.6387912 · Cited by 115 · see citing works
Real-time reaction monitoring by continuous-introduction ion-spray tandem mass spectrometry
Lee, E. D., Mueck, W., Henion, J. D., et al. (1989) · Journal of the American Chemical Society · doi:10.1021/ja00195a012 · Cited by 107 · see citing works
Self-citationCapillary zone electrophoresis/tandem mass spectrometry for the determination of sulfonated azo dyes
Lee, E. D., Mück, W., Henion, J. D., et al. (1989) · Journal of Mass Spectrometry · doi:10.1002/bms.1200180408 · Cited by 105 · see citing works
Self-citationLC/MS and LC/MS/MS determination of protein tryptic digests
Huang, E. C., & Henion, J. D. (1990) · Journal of the American Society for Mass Spectrometry · doi:10.1016/1044-0305(90)85052-n · Cited by 104 · see citing works
Self-citationHuang, E. C., & Henion, J. D. (1991) · Analytical Chemistry · doi:10.1021/ac00007a016 · Cited by 77 · see citing works
Self-citationCarr, S. A., Hemling, M. E., Folena-Wasserman, G., et al. (1989) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)30077-8 · Cited by 72 · see citing works
Henzel, W. J., Bourell, J. H., & Stults, J. T. (1990) · Analytical Biochemistry · doi:10.1016/0003-2697(90)90448-i · Cited by 65 · see citing works
Siu, K. W. M., Gardner, G. J., & Berman, S. S. (1989) · Analytical Chemistry · doi:10.1021/ac00195a021 · Cited by 64 · see citing works
Mück, W. M., & Henion, J. D. (1989) · Journal of Chromatography B Biomedical Sciences and Applications · doi:10.1016/s0378-4347(00)82608-0 · Cited by 58 · see citing works
Self-citationCaprioli, R. M., DaGue, B. B., Fan, T., et al. (1987) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(87)90724-8 · Cited by 55 · see citing works
Electrospray ionization on a high‐performance magnetic‐sector mass spectrometer
Meng, C., McEwen, C. N., & Larsen, B. S. (1990) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290040504 · Cited by 52 · see citing works
Oligopeptide sequence analysis by collision‐activated dissociation of multiply charged ions
Hunt, D. F., Zhu, N., Shabanowitz, J., et al. (1989) · Rapid Communications in Mass Spectrometry · doi:10.1002/rcm.1290030408 · Cited by 47 · see citing works
Caprioli, R. M., Moore, W. T., DaGue, B. B., et al. (1988) · Journal of Chromatography A · doi:10.1016/s0021-9673(00)94807-6 · Cited by 47 · see citing works
Lee, E. D., Henion, J. D., & Covey, T. R. (1989) · Journal of Microcolumn Separations · doi:10.1002/mcs.1220010105 · Cited by 42 · see citing works
Self-citationRapid protein sequencing by the enzyme-thermospray liquid chromatographic/mass spectrometric method
Stachowiak, K., Wilder, C., Vestal, M. L., et al. (1988) · Journal of the American Chemical Society · doi:10.1021/ja00214a017 · Cited by 36 · see citing works
Analysis of middle mass peptides by thermospray LC/MS
Rudewicz, P. J. (1988) · Journal of Mass Spectrometry · doi:10.1002/bms.1200150809 · Cited by 12 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Proteomics by Mass Spectrometry: Approaches, Advances, and Applications
Yates, J. R., Ruse, C. I., & Nakorchevsky, A. (2009) · Annual Review of Biomedical Engineering · doi:10.1146/annurev-bioeng-061008-124934 · Cited by 1,078 · see citing works
Quantitative, High-Resolution Proteomics for Data-Driven Systems Biology
Cox, J., & Mann, M. (2011) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-061308-093216 · Cited by 685 · see citing works
Juillard, V, Laan, H., Kunji, E. R., Jeronimus-Stratingh, C. M., Bruins, A. P., & Konings, W. N. (1995) · Journal of Bacteriology · doi:10.1128/jb.177.12.3472-3478.1995 · Cited by 156 · see citing works
- Authors
- Covey, T. R., Huang, E. C., & Henion, J. D.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1991-07-01
- Volume
- 63
- Issue
- 13
- Pages
- 1193-1200
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W1988287443
- Cited by
- 446
- References
- 43
- Citations by year
- 2025: 2 · 2023: 3 · 2022: 1 · 2021: 3 · 2020: 1 · 2019: 2 · 2018: 4 · 2017: 2 · 2016: 6 · 2015: 5 · 2014: 5 · 2013: 9 · 2012: 7
- FWCI
- 24.13
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Keywords
- tryptic peptides, collision-induced dissociation, electrospray ionization, molecular weight determination, human hemoglobin, peptide structural characterization
- MeSH terms
- Amino Acid Sequence, Chromatography, Liquid, Hemoglobins, Molecular Sequence Data, Peptide Fragments, Growth Hormone, Mass Spectrometry, Trypsin, Human Growth Hormone
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
