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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of 4-sulfophenyl isothiocyanate-derivatized peptides on AnchorChip sample supports using the sodium-tolerant matrix 2,4,6-trihydroxyacetophenone and diammonium citrate.
Rapid Commun Mass Spectrom. 2005; 19(6):752-8.RC

Abstract

The reagent 4-sulfophenyl isothiocyanate (SPITC) is an effective, stable, and inexpensive alternative to commercially available reagents used in the N-terminal sulfonation of peptides for enhanced postsource decay (PSD) in matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOFMS) analyses. However, suppression of ionization of sulfonated peptides due to sample and matrix contaminants such as sodium can be a problem when using prestructured MALDI target sample supports, such as the Bruker Daltonics AnchorChip. We show that use of the salt-tolerant matrix 2,4,6-trihydroxyacetophenone containing diammonium citrate (THAP/DAC) as an alternative to alpha-cyanohydroxycinnamic acid (HCCA) reduces the need for extensive washing of ZipTip-bound peptides or additional on-target sample clean-up steps. Use of the THAP/DAC matrix results in selective ionization of sulfonated peptides with greater peptide coverage, as well as detection of higher mass derivatized peptides, than was observed for HCCA or THAP alone. The THAP/DAC matrix is quite tolerant of sodium contamination, with SPITC-peptides detectable in preparations containing up to 50 mM NaCl. In addition, THAP/DAC matrix was found to promote efficient PSD fragmentation of sulfonated peptides. We demonstrated the utility of using the THAP/DAC MALDI matrix for peptide sequencing with DNA polymerase beta tryptic peptide mixture, as well as tryptic peptides derived from Xiphophorus maculatus brain extract proteins previously separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE).

Authors+Show Affiliations

Oehlers LP
Molecular Biosciences Research Group, Department of Chemistry and Biochemistry, Texas State University, San Marcos, TX 78666, USA.
Perez AN
No affiliation info available
Walter RB
No affiliation info available

MeSH

AcetophenonesAnimalsBrainCyprinodontiformesDNA Polymerase betaDrug ToleranceIsothiocyanatesMenNeuropeptidesProtein Array AnalysisSequence Analysis, ProteinSodiumSpectrometry, Mass, Electrospray IonizationSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Pub Type(s)

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

Language

eng

PubMed ID

15712288

Citation

Oehlers, Leon P., et al. "Matrix-assisted Laser Desorption/ionization Time-of-flight Mass Spectrometry of 4-sulfophenyl Isothiocyanate-derivatized Peptides On AnchorChip Sample Supports Using the Sodium-tolerant Matrix 2,4,6-trihydroxyacetophenone and Diammonium Citrate." Rapid Communications in Mass Spectrometry : RCM, vol. 19, no. 6, 2005, pp. 752-8.
Oehlers LP, Perez AN, Walter RB. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of 4-sulfophenyl isothiocyanate-derivatized peptides on AnchorChip sample supports using the sodium-tolerant matrix 2,4,6-trihydroxyacetophenone and diammonium citrate. Rapid Commun Mass Spectrom. 2005;19(6):752-8.
Oehlers, L. P., Perez, A. N., & Walter, R. B. (2005). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of 4-sulfophenyl isothiocyanate-derivatized peptides on AnchorChip sample supports using the sodium-tolerant matrix 2,4,6-trihydroxyacetophenone and diammonium citrate. Rapid Communications in Mass Spectrometry : RCM, 19(6), 752-8.
Oehlers LP, Perez AN, Walter RB. Matrix-assisted Laser Desorption/ionization Time-of-flight Mass Spectrometry of 4-sulfophenyl Isothiocyanate-derivatized Peptides On AnchorChip Sample Supports Using the Sodium-tolerant Matrix 2,4,6-trihydroxyacetophenone and Diammonium Citrate. Rapid Commun Mass Spectrom. 2005;19(6):752-8. PubMed PMID: 15712288.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of 4-sulfophenyl isothiocyanate-derivatized peptides on AnchorChip sample supports using the sodium-tolerant matrix 2,4,6-trihydroxyacetophenone and diammonium citrate. AU - Oehlers,Leon P, AU - Perez,Amy N, AU - Walter,Ronald B, PY - 2005/2/16/pubmed PY - 2005/5/13/medline PY - 2005/2/16/entrez SP - 752 EP - 8 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 19 IS - 6 N2 - The reagent 4-sulfophenyl isothiocyanate (SPITC) is an effective, stable, and inexpensive alternative to commercially available reagents used in the N-terminal sulfonation of peptides for enhanced postsource decay (PSD) in matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOFMS) analyses. However, suppression of ionization of sulfonated peptides due to sample and matrix contaminants such as sodium can be a problem when using prestructured MALDI target sample supports, such as the Bruker Daltonics AnchorChip. We show that use of the salt-tolerant matrix 2,4,6-trihydroxyacetophenone containing diammonium citrate (THAP/DAC) as an alternative to alpha-cyanohydroxycinnamic acid (HCCA) reduces the need for extensive washing of ZipTip-bound peptides or additional on-target sample clean-up steps. Use of the THAP/DAC matrix results in selective ionization of sulfonated peptides with greater peptide coverage, as well as detection of higher mass derivatized peptides, than was observed for HCCA or THAP alone. The THAP/DAC matrix is quite tolerant of sodium contamination, with SPITC-peptides detectable in preparations containing up to 50 mM NaCl. In addition, THAP/DAC matrix was found to promote efficient PSD fragmentation of sulfonated peptides. We demonstrated the utility of using the THAP/DAC MALDI matrix for peptide sequencing with DNA polymerase beta tryptic peptide mixture, as well as tryptic peptides derived from Xiphophorus maculatus brain extract proteins previously separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/15712288/Matrix_assisted_laser_desorption/ionization_time_of_flight_mass_spectrometry_of_4_sulfophenyl_isothiocyanate_derivatized_peptides_on_AnchorChip_sample_supports_using_the_sodium_tolerant_matrix_246_trihydroxyacetophenone_and_diammonium_citrate_ DB - PRIME DP - Unbound Medicine ER -
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