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De novo sequencing of tryptic peptides sulfonated by 4-sulfophenyl isothiocyanate for unambiguous protein identification using post-source decay matrix-assisted laser desorption/ionization mass spectrometry.
Rapid Commun Mass Spectrom. 2004; 18(2):191-8.RC

Abstract

A simple method of solid-phase derivatization and sequencing of tryptic peptides has been developed for rapid and unambiguous identification of spots on two-dimensional gels using post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The proteolytic digests of proteins are chemically modified by 4-sulfophenyl isothiocyanate. The derivatization reaction introduces a negative sulfonic acid group at the N-terminus of a peptide, which can increase the efficiency of PSD fragmentation and enable the selective detection of only a single series of fragment ions (y-ions). This chemically assisted method avoids the limitation of high background normally observed in MALDI-PSD spectra, and makes the spectra easier to interpret and facilitates de novo sequencing of internal fragment. The modification reaction is conducted in C(18) microZipTips to decrease the background and to enhance the signal/noise. Derivatization procedures were optimized for MALDI-PSD to increase the structural information and to obtain a complete peptide sequence even in critical cases. The MALDI-PSD mass spectra of two model peptides and their sulfonated derivatives are compared. For some proteins unambiguous identification could be achieved by MALDI-PSD sequencing of derivatized peptides obtained from in-gel digests of phosphorylase B and proteins of hepatic satellite cells (HSC).

Authors+Show Affiliations

College of Life Sciences, Hunan Normal University, Changsha, 410081 China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

14745769

Citation

Chen, Ping, et al. "De Novo Sequencing of Tryptic Peptides Sulfonated By 4-sulfophenyl Isothiocyanate for Unambiguous Protein Identification Using Post-source Decay Matrix-assisted Laser Desorption/ionization Mass Spectrometry." Rapid Communications in Mass Spectrometry : RCM, vol. 18, no. 2, 2004, pp. 191-8.
Chen P, Nie S, Mi W, et al. De novo sequencing of tryptic peptides sulfonated by 4-sulfophenyl isothiocyanate for unambiguous protein identification using post-source decay matrix-assisted laser desorption/ionization mass spectrometry. Rapid Commun Mass Spectrom. 2004;18(2):191-8.
Chen, P., Nie, S., Mi, W., Wang, X. C., & Liang, S. P. (2004). De novo sequencing of tryptic peptides sulfonated by 4-sulfophenyl isothiocyanate for unambiguous protein identification using post-source decay matrix-assisted laser desorption/ionization mass spectrometry. Rapid Communications in Mass Spectrometry : RCM, 18(2), 191-8.
Chen P, et al. De Novo Sequencing of Tryptic Peptides Sulfonated By 4-sulfophenyl Isothiocyanate for Unambiguous Protein Identification Using Post-source Decay Matrix-assisted Laser Desorption/ionization Mass Spectrometry. Rapid Commun Mass Spectrom. 2004;18(2):191-8. PubMed PMID: 14745769.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - De novo sequencing of tryptic peptides sulfonated by 4-sulfophenyl isothiocyanate for unambiguous protein identification using post-source decay matrix-assisted laser desorption/ionization mass spectrometry. AU - Chen,Ping, AU - Nie,Song, AU - Mi,Wei, AU - Wang,Xian-Chun, AU - Liang,Song-Ping, PY - 2004/1/28/pubmed PY - 2004/3/27/medline PY - 2004/1/28/entrez SP - 191 EP - 8 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 18 IS - 2 N2 - A simple method of solid-phase derivatization and sequencing of tryptic peptides has been developed for rapid and unambiguous identification of spots on two-dimensional gels using post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The proteolytic digests of proteins are chemically modified by 4-sulfophenyl isothiocyanate. The derivatization reaction introduces a negative sulfonic acid group at the N-terminus of a peptide, which can increase the efficiency of PSD fragmentation and enable the selective detection of only a single series of fragment ions (y-ions). This chemically assisted method avoids the limitation of high background normally observed in MALDI-PSD spectra, and makes the spectra easier to interpret and facilitates de novo sequencing of internal fragment. The modification reaction is conducted in C(18) microZipTips to decrease the background and to enhance the signal/noise. Derivatization procedures were optimized for MALDI-PSD to increase the structural information and to obtain a complete peptide sequence even in critical cases. The MALDI-PSD mass spectra of two model peptides and their sulfonated derivatives are compared. For some proteins unambiguous identification could be achieved by MALDI-PSD sequencing of derivatized peptides obtained from in-gel digests of phosphorylase B and proteins of hepatic satellite cells (HSC). SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/14745769/De_novo_sequencing_of_tryptic_peptides_sulfonated_by_4_sulfophenyl_isothiocyanate_for_unambiguous_protein_identification_using_post_source_decay_matrix_assisted_laser_desorption/ionization_mass_spectrometry_ L2 - https://doi.org/10.1002/rcm.1280 DB - PRIME DP - Unbound Medicine ER -