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Identification of proteins from two-dimensional gel electrophoresis of human erythroleukemia cells using capillary high performance liquid chromatography/electrospray-ion trap-reflectron time-of-flight mass spectrometry with two-dimensional topographic map analysis of in-gel tryptic digest products.
Rapid Commun Mass Spectrom. 1999; 13(19):1907-16.RC

Abstract

Protein spots from two-dimensional (2-D) gel electrophoresis of a human erythroleukemia cell line have been identified by analysis of the in-gel tryptic digests using capillary high performance liquid chromatography (HPLC) separation with on-line detection using electrospray ionization mass spectrometry (ESI-MS). This is performed using an electrospray/ion trap storage/reflectron time-of-flight mass spectrometer system (ESI-IT-reTOFMS). A 2-D topographic mapping display developed to process the on-line data acquired with this TOF system has been used to obtain mass identification of each peptide, even though the capillary HPLC only provides limited separation capability of the tryptic peptide mixtures studied herein. Using this method, a substantial fraction of the protein sequence can be covered and identified using the tryptic map. It is demonstrated that by entering the cell species, the approximate MW and pI range as determined by 2-D gel electrophoresis, and the tryptic peptide map into the database a unique match for identification of the protein generally results. It is also demonstrated that a much improved coverage of the protein sequence is obtained by this method relative to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS).

Authors+Show Affiliations

Department of Chemistry, School of Medicine, The University of Michigan, Ann Arbor, MI 48109-1055, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10487937

Citation

Chen, Y, et al. "Identification of Proteins From Two-dimensional Gel Electrophoresis of Human Erythroleukemia Cells Using Capillary High Performance Liquid Chromatography/electrospray-ion Trap-reflectron Time-of-flight Mass Spectrometry With Two-dimensional Topographic Map Analysis of In-gel Tryptic Digest Products." Rapid Communications in Mass Spectrometry : RCM, vol. 13, no. 19, 1999, pp. 1907-16.
Chen Y, Jin X, Misek D, et al. Identification of proteins from two-dimensional gel electrophoresis of human erythroleukemia cells using capillary high performance liquid chromatography/electrospray-ion trap-reflectron time-of-flight mass spectrometry with two-dimensional topographic map analysis of in-gel tryptic digest products. Rapid Commun Mass Spectrom. 1999;13(19):1907-16.
Chen, Y., Jin, X., Misek, D., Hinderer, R., Hanash, S. M., & Lubman, D. M. (1999). Identification of proteins from two-dimensional gel electrophoresis of human erythroleukemia cells using capillary high performance liquid chromatography/electrospray-ion trap-reflectron time-of-flight mass spectrometry with two-dimensional topographic map analysis of in-gel tryptic digest products. Rapid Communications in Mass Spectrometry : RCM, 13(19), 1907-16.
Chen Y, et al. Identification of Proteins From Two-dimensional Gel Electrophoresis of Human Erythroleukemia Cells Using Capillary High Performance Liquid Chromatography/electrospray-ion Trap-reflectron Time-of-flight Mass Spectrometry With Two-dimensional Topographic Map Analysis of In-gel Tryptic Digest Products. Rapid Commun Mass Spectrom. 1999;13(19):1907-16. PubMed PMID: 10487937.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of proteins from two-dimensional gel electrophoresis of human erythroleukemia cells using capillary high performance liquid chromatography/electrospray-ion trap-reflectron time-of-flight mass spectrometry with two-dimensional topographic map analysis of in-gel tryptic digest products. AU - Chen,Y, AU - Jin,X, AU - Misek,D, AU - Hinderer,R, AU - Hanash,S M, AU - Lubman,D M, PY - 1999/9/17/pubmed PY - 1999/9/17/medline PY - 1999/9/17/entrez SP - 1907 EP - 16 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 13 IS - 19 N2 - Protein spots from two-dimensional (2-D) gel electrophoresis of a human erythroleukemia cell line have been identified by analysis of the in-gel tryptic digests using capillary high performance liquid chromatography (HPLC) separation with on-line detection using electrospray ionization mass spectrometry (ESI-MS). This is performed using an electrospray/ion trap storage/reflectron time-of-flight mass spectrometer system (ESI-IT-reTOFMS). A 2-D topographic mapping display developed to process the on-line data acquired with this TOF system has been used to obtain mass identification of each peptide, even though the capillary HPLC only provides limited separation capability of the tryptic peptide mixtures studied herein. Using this method, a substantial fraction of the protein sequence can be covered and identified using the tryptic map. It is demonstrated that by entering the cell species, the approximate MW and pI range as determined by 2-D gel electrophoresis, and the tryptic peptide map into the database a unique match for identification of the protein generally results. It is also demonstrated that a much improved coverage of the protein sequence is obtained by this method relative to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/10487937/Identification_of_proteins_from_two_dimensional_gel_electrophoresis_of_human_erythroleukemia_cells_using_capillary_high_performance_liquid_chromatography/electrospray_ion_trap_reflectron_time_of_flight_mass_spectrometry_with_two_dimensional_topographic_map_analysis_of_in_gel_tryptic_digest_products_ DB - PRIME DP - Unbound Medicine ER -