Details

Autor(en) / Beteiligte

• Horvatić, Anita
• Dodig, Ivana
• Vuletić, Tomislav
• Pavoković, Dubravko
• Hameršak, Zdenko
• Butorac, Ana
• Cindrić, Mario

Titel

Comparison between Enhanced MALDI In-source Decay by Ammonium Persulfate and N- or C‑Terminal Derivatization Methods for Detailed Peptide Structure Determination

Ist Teil von

• Analytical chemistry (Washington), 2013-04, Vol.85 (8), p.3940-3947

Ort / Verlag

United States: American Chemical Society

Erscheinungsjahr

2013

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Amino acid sequencing and more detailed structure elucidation analysis of peptides and small proteins is a very difficult task even if state-of-the-art mass spectrometry (MS) is employed. To make this task easier, chemical derivatization methods of the N terminus with 4-sulfophenyl-isothiocyanate (SPITC) or the C terminus with 2-methoxy-4,5-dihydro-1H-imidazole (Lys-tag) can enhance peptide fragmentation or fragment ionizability, via proton mobility/localization mechanisms making tandem MS (MS2) spectra more informative and less demanding for structural interpretation. Observed disadvantages related to both derivatization methods are sample- and time-consuming procedures and the increased number of reaction byproducts. A novel, sulfate radical in-source formation method of matrix-assisted laser desorption ionization (MALDI) MS based on chemically enhanced in-source decay (ISD) can be accomplished by simple addition of ammonium persulfate (APS) in the matrix solution. This method enables effective decomposition of peptide ions already in the first stage of MS analysis where a large number of fragment ions are produced. The resultant MALDI-ISD mass spectra (MS after APS → MALDI-ISD MS) are almost equivalent to conventional, collision-induced dissociation (CID) MS2 spectra. These fragment ions are further subjected to the second stage of the MS, and consequently, MS3 spectra are produced, which makes the sequence analysis more informative and complete (CID MS2 is thus equivalent to CID MS3). Multiply stage MS after APS addition showed enhanced sensitivity, resolution, and mass accuracy compared to peptide derivatization (SPITC and Lys-tag) or conventional MS and MS2 analyses and offered more detailed insight into peptide structure.