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Easy amino acid sequencing of sulfonated peptides using post-source decay on a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer equipped with a variable voltage reflector
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Ludwig Institute for Cancer Research.
2002 (English)In: Rapid Communications in Mass Spectrometry, ISSN 0951-4198, E-ISSN 1097-0231, Vol. 16, no 19, 1851-1859 p.Article in journal (Refereed) Published
Abstract [en]

Tryptic peptides were labeled with sulfonic acid groups at the N-termini using an improved chemistry. The derivatization was performed in common aqueous buffers on peptides adsorbed onto a ZipTip trade mark C(18), thus allowing simultaneous desalting/concentration of the sample. When only Arg-terminating peptides were considered, the procedure from adsorption onto the ZipTip until analysis by MALDI-PSD took about 10 min and several samples could be worked on in parallel. The resulting improved post-source decay (PSD) fragmentation produced spectra containing only y-ions. PSD amino acid sequencing of underivatized and derivatized synthetic peptides was compared. From the sequence information obtained from derivatized peptides isolated by ion selection from tryptic in-gel digests, a protein was correctly identified which was difficult to analyze from an unclear peptide mass fingerprint analysis. The method was also applied to the identification and localization of phosphorylated Ser and Tyr residues in native and synthetic peptides.

Place, publisher, year, edition, pages
2002. Vol. 16, no 19, 1851-1859 p.
Keyword [en]
Guanidines/chemistry, Hydrolysis, Indicators and Reagents, Lysine/chemistry, Peptides/*analysis, Phosphopeptides/analysis, Sequence Analysis, Spectrometry; Mass; Matrix-Assisted Laser Desorption-Ionization, Sulfonic Acids/analysis, Trypsin
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-10408DOI: 10.1002/rcm.805PubMedID: 12271450OAI: oai:DiVA.org:uu-10408DiVA: diva2:38176
Available from: 2007-03-22 Created: 2007-03-22 Last updated: 2017-12-11Bibliographically approved

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Hellman, Ulf

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