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Matrix-assisted laser desorption/ionization directed nano-electrospray ionization tandem mass spectrometric analysis for protein identification.
Rapid Commun Mass Spectrom. 2003; 17(16):1825-34.RC

Abstract

In those cases where the information obtained by peptide mass fingerprinting or matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-MS/MS) is not sufficient for unambiguous protein identification, nano-electrospray ionization (nano-ESI) and/or electrospray ionization tandem mass spectrometry (ESI-MS/MS) analysis must be performed. The sensitivity of nano-ESI/MS, however, is lower than that of MALDI-MS, especially at very low analyte concentrations and/or in the presence of contaminants, such as salt and detergents. Moreover, to perform ESI-MS/MS, the peptide masses of the precursor ions must be known. The approach described in this paper, MALDI-directed nano-ESI-MS/MS, makes use of information obtained from the more sensitive MALDI-MS experiments in order to direct subsequent nano-ESI-MS/MS experiments. Peptide molecular ions found in the MALDI-MS analysis are then selected, as their (+2) precursor ions, for nano-ESI-MS/MS sequencing, even though these ions cannot be detected in the ESI-MS spectra. This method, originally proposed by Tempst et al. (Anal. Chem. 2000, 72: 777-790), has been extended to provide better sensitivity and shorter analysis times; also, a comparison with liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been performed. These experiments, performed using quadrupole time-of-flight instruments equipped with commercially available nano-ESI sources, have allowed the unambiguous identification of in-gel digested proteins at levels below their ESI-MS detection limits, even in the presence of salts and detergents.

Authors+Show Affiliations

Kast J
Biomedical Research Centre, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada.
Parker CE
No affiliation info available
van der Drift K
No affiliation info available
Dial JM
No affiliation info available
Milgram SL
No affiliation info available
Wilm M
No affiliation info available
Howell M
No affiliation info available
Borchers CH
No affiliation info available

MeSH

NanotechnologyProteinsSensitivity and SpecificitySpectrometry, Mass, Electrospray IonizationSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Pub Type(s)

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

Language

eng

PubMed ID

12876682

Citation

Kast, Juergen, et al. "Matrix-assisted Laser Desorption/ionization Directed Nano-electrospray Ionization Tandem Mass Spectrometric Analysis for Protein Identification." Rapid Communications in Mass Spectrometry : RCM, vol. 17, no. 16, 2003, pp. 1825-34.
Kast J, Parker CE, van der Drift K, et al. Matrix-assisted laser desorption/ionization directed nano-electrospray ionization tandem mass spectrometric analysis for protein identification. Rapid Commun Mass Spectrom. 2003;17(16):1825-34.
Kast, J., Parker, C. E., van der Drift, K., Dial, J. M., Milgram, S. L., Wilm, M., Howell, M., & Borchers, C. H. (2003). Matrix-assisted laser desorption/ionization directed nano-electrospray ionization tandem mass spectrometric analysis for protein identification. Rapid Communications in Mass Spectrometry : RCM, 17(16), 1825-34.
Kast J, et al. Matrix-assisted Laser Desorption/ionization Directed Nano-electrospray Ionization Tandem Mass Spectrometric Analysis for Protein Identification. Rapid Commun Mass Spectrom. 2003;17(16):1825-34. PubMed PMID: 12876682.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Matrix-assisted laser desorption/ionization directed nano-electrospray ionization tandem mass spectrometric analysis for protein identification. AU - Kast,Juergen, AU - Parker,Carol E, AU - van der Drift,Koen, AU - Dial,J Michael, AU - Milgram,Sharon L, AU - Wilm,Matthias, AU - Howell,Michael, AU - Borchers,Christoph H, PY - 2003/7/24/pubmed PY - 2003/9/23/medline PY - 2003/7/24/entrez SP - 1825 EP - 34 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 17 IS - 16 N2 - In those cases where the information obtained by peptide mass fingerprinting or matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI-MS/MS) is not sufficient for unambiguous protein identification, nano-electrospray ionization (nano-ESI) and/or electrospray ionization tandem mass spectrometry (ESI-MS/MS) analysis must be performed. The sensitivity of nano-ESI/MS, however, is lower than that of MALDI-MS, especially at very low analyte concentrations and/or in the presence of contaminants, such as salt and detergents. Moreover, to perform ESI-MS/MS, the peptide masses of the precursor ions must be known. The approach described in this paper, MALDI-directed nano-ESI-MS/MS, makes use of information obtained from the more sensitive MALDI-MS experiments in order to direct subsequent nano-ESI-MS/MS experiments. Peptide molecular ions found in the MALDI-MS analysis are then selected, as their (+2) precursor ions, for nano-ESI-MS/MS sequencing, even though these ions cannot be detected in the ESI-MS spectra. This method, originally proposed by Tempst et al. (Anal. Chem. 2000, 72: 777-790), has been extended to provide better sensitivity and shorter analysis times; also, a comparison with liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been performed. These experiments, performed using quadrupole time-of-flight instruments equipped with commercially available nano-ESI sources, have allowed the unambiguous identification of in-gel digested proteins at levels below their ESI-MS detection limits, even in the presence of salts and detergents. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/12876682/Matrix_assisted_laser_desorption/ionization_directed_nano_electrospray_ionization_tandem_mass_spectrometric_analysis_for_protein_identification_ DB - PRIME DP - Unbound Medicine ER -
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