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Derivatization procedures to facilitate de novo sequencing of lysine-terminated tryptic peptides using postsource decay matrix-assisted laser desorption/ionization mass spectrometry.
Rapid Commun Mass Spectrom. 2000; 14(24):2348-56.RC

Abstract

Guanidination of the epsilon-amino group of lysine-terminated tryptic peptides can be accomplished selectively in one step with O-methylisourea hydrogen sulfate. This reaction converts lysine residues into more basic homoarginine residues. It also protects the epsilon-amino groups against unwanted reaction with sulfonation reagents, which can then be used to selectively modify the N-termini of tryptic peptides. The combined reactions convert lysine-terminated tryptic peptides into modified peptides that are suitable for de novo sequencing by postsource decay matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The guanidination reaction is very pH dependent. Product yields and reaction kinetics were studied in aqueous solution using either NaOH or diisopropylethylamine as the base. Methods are reported for derivatizing and sequencing lysine-terminated tryptic peptides at low pmole levels. The postsource decay (PSD) MALDI tandem mass spectra of a model peptide (VGGYGYGAK), the homoarginine analog and the sulfonated homoarginine analog are compared. These spectra show the influence that each chemical modification has on the peptide fragmentation pattern. Finally, we demonstrate that definitive protein identifications can be achieved by PSD MALDI sequencing of derivatized peptides obtained from solution digests of model proteins and from in-gel digests of 2D-gel separated proteins.

Authors+Show Affiliations

The Procter and Gamble Co., Miami Valley Laboratories, PO Box 538707, Cincinnati, OH 45253-8707, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11114049

Citation

Keough, T, et al. "Derivatization Procedures to Facilitate De Novo Sequencing of Lysine-terminated Tryptic Peptides Using Postsource Decay Matrix-assisted Laser Desorption/ionization Mass Spectrometry." Rapid Communications in Mass Spectrometry : RCM, vol. 14, no. 24, 2000, pp. 2348-56.
Keough T, Lacey MP, Youngquist RS. Derivatization procedures to facilitate de novo sequencing of lysine-terminated tryptic peptides using postsource decay matrix-assisted laser desorption/ionization mass spectrometry. Rapid Commun Mass Spectrom. 2000;14(24):2348-56.
Keough, T., Lacey, M. P., & Youngquist, R. S. (2000). Derivatization procedures to facilitate de novo sequencing of lysine-terminated tryptic peptides using postsource decay matrix-assisted laser desorption/ionization mass spectrometry. Rapid Communications in Mass Spectrometry : RCM, 14(24), 2348-56.
Keough T, Lacey MP, Youngquist RS. Derivatization Procedures to Facilitate De Novo Sequencing of Lysine-terminated Tryptic Peptides Using Postsource Decay Matrix-assisted Laser Desorption/ionization Mass Spectrometry. Rapid Commun Mass Spectrom. 2000;14(24):2348-56. PubMed PMID: 11114049.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Derivatization procedures to facilitate de novo sequencing of lysine-terminated tryptic peptides using postsource decay matrix-assisted laser desorption/ionization mass spectrometry. AU - Keough,T, AU - Lacey,M P, AU - Youngquist,R S, PY - 2000/12/13/pubmed PY - 2001/3/3/medline PY - 2000/12/13/entrez SP - 2348 EP - 56 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 14 IS - 24 N2 - Guanidination of the epsilon-amino group of lysine-terminated tryptic peptides can be accomplished selectively in one step with O-methylisourea hydrogen sulfate. This reaction converts lysine residues into more basic homoarginine residues. It also protects the epsilon-amino groups against unwanted reaction with sulfonation reagents, which can then be used to selectively modify the N-termini of tryptic peptides. The combined reactions convert lysine-terminated tryptic peptides into modified peptides that are suitable for de novo sequencing by postsource decay matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The guanidination reaction is very pH dependent. Product yields and reaction kinetics were studied in aqueous solution using either NaOH or diisopropylethylamine as the base. Methods are reported for derivatizing and sequencing lysine-terminated tryptic peptides at low pmole levels. The postsource decay (PSD) MALDI tandem mass spectra of a model peptide (VGGYGYGAK), the homoarginine analog and the sulfonated homoarginine analog are compared. These spectra show the influence that each chemical modification has on the peptide fragmentation pattern. Finally, we demonstrate that definitive protein identifications can be achieved by PSD MALDI sequencing of derivatized peptides obtained from solution digests of model proteins and from in-gel digests of 2D-gel separated proteins. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/11114049/Derivatization_procedures_to_facilitate_de_novo_sequencing_of_lysine_terminated_tryptic_peptides_using_postsource_decay_matrix_assisted_laser_desorption/ionization_mass_spectrometry_ L2 - https://doi.org/10.1002/1097-0231(20001230)14:24<2348::AID-RCM175>3.0.CO;2-8 DB - PRIME DP - Unbound Medicine ER -