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Comparative identification of prostanoid inducible proteins by LC-ESI-MS/MS and MALDI-TOF mass spectrometry.
Chem Res Toxicol. 2003 Jun; 16(6):757-67.CR

Abstract

Protein identification by MS is well-established. Mixtures of proteins from cell extracts are separated by either one- or two-dimensional gel electrophoresis, and specific bands or spots are subjected to in-gel digestion and subsequent analysis by MS. The two most common types of ionization used in MS are electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI). When ESI is used, the sample is typically analyzed by inline HPLC-ESI-MS/MS with fragmentation of individual digest peptides, followed by database comparison between theoretical and experimental fragmentation patterns. MALDI-MS analysis is based on peptide mass mapping, with mass measurements of the digest peptides searched against a database of theoretical digests. We give here the results of a comparison between ESI-ion trap and MALDI-TOF (time-of-flight) analysis of 11-deoxy,16,16-dimethyl prostaglandin E(2) (DDM-PGE(2)) inducible proteins. Individual peptides identified by the two techniques differed, in general, but the resulting protein identification was the same. Slightly higher coverage of each protein was obtained by MALDI-TOF, but the MS/MS data were more definitive by requiring fewer peptides to assign a positive identification. Both methods effectively identified two proteins in the same gel band. The samples here are derived from a renal epithelial cell line (LLC-PK(1)) established from the New Hampshire minipig, a species poorly represented in the current database, and strategies and limitations for analyzing such species are discussed.

Authors+Show Affiliations

Center for Molecular and Cellular Toxicology, Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas at Austin, Austin, Texas 78712, 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, P.H.S.

Language

eng

PubMed ID

12807359

Citation

Person, Maria D., et al. "Comparative Identification of Prostanoid Inducible Proteins By LC-ESI-MS/MS and MALDI-TOF Mass Spectrometry." Chemical Research in Toxicology, vol. 16, no. 6, 2003, pp. 757-67.
Person MD, Lo HH, Towndrow KM, et al. Comparative identification of prostanoid inducible proteins by LC-ESI-MS/MS and MALDI-TOF mass spectrometry. Chem Res Toxicol. 2003;16(6):757-67.
Person, M. D., Lo, H. H., Towndrow, K. M., Jia, Z., Monks, T. J., & Lau, S. S. (2003). Comparative identification of prostanoid inducible proteins by LC-ESI-MS/MS and MALDI-TOF mass spectrometry. Chemical Research in Toxicology, 16(6), 757-67.
Person MD, et al. Comparative Identification of Prostanoid Inducible Proteins By LC-ESI-MS/MS and MALDI-TOF Mass Spectrometry. Chem Res Toxicol. 2003;16(6):757-67. PubMed PMID: 12807359.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparative identification of prostanoid inducible proteins by LC-ESI-MS/MS and MALDI-TOF mass spectrometry. AU - Person,Maria D, AU - Lo,Herng-Hsiang, AU - Towndrow,Kelly M, AU - Jia,Zhe, AU - Monks,Terrence J, AU - Lau,Serrine S, PY - 2003/6/17/pubmed PY - 2004/2/5/medline PY - 2003/6/17/entrez SP - 757 EP - 67 JF - Chemical research in toxicology JO - Chem Res Toxicol VL - 16 IS - 6 N2 - Protein identification by MS is well-established. Mixtures of proteins from cell extracts are separated by either one- or two-dimensional gel electrophoresis, and specific bands or spots are subjected to in-gel digestion and subsequent analysis by MS. The two most common types of ionization used in MS are electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI). When ESI is used, the sample is typically analyzed by inline HPLC-ESI-MS/MS with fragmentation of individual digest peptides, followed by database comparison between theoretical and experimental fragmentation patterns. MALDI-MS analysis is based on peptide mass mapping, with mass measurements of the digest peptides searched against a database of theoretical digests. We give here the results of a comparison between ESI-ion trap and MALDI-TOF (time-of-flight) analysis of 11-deoxy,16,16-dimethyl prostaglandin E(2) (DDM-PGE(2)) inducible proteins. Individual peptides identified by the two techniques differed, in general, but the resulting protein identification was the same. Slightly higher coverage of each protein was obtained by MALDI-TOF, but the MS/MS data were more definitive by requiring fewer peptides to assign a positive identification. Both methods effectively identified two proteins in the same gel band. The samples here are derived from a renal epithelial cell line (LLC-PK(1)) established from the New Hampshire minipig, a species poorly represented in the current database, and strategies and limitations for analyzing such species are discussed. SN - 0893-228X UR - https://www.unboundmedicine.com/medline/citation/12807359/Comparative_identification_of_prostanoid_inducible_proteins_by_LC_ESI_MS/MS_and_MALDI_TOF_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -