Use of a novel ionic liquid matrix for MALDI-MS analysis of glycopeptides and glycans out of total tryptic digests.
In this study, we investigated a novel ionic liquid matrix (ILM), namely, the 1,1,3,3-tetramethylguanidinium salt of 2,4,6-trihydroxyacetophenone (THAP). This matrix[1,1,3,3-tetramethylguanidinium 2,4,6-trihydroxyacetophenone (GTHAP)] turned out to be well suited for the matrix-assisted laser desorption/ionization mass spectrometric analysis of glycopeptides and glycans, and overcame the well-known ionization suppression of carbohydrate structures in the presence of peptides. The matrix was evaluated by two different series of experiments, in each case in comparison with the crystalline THAP matrix. In the first set of experiments, mass spectra were taken from unseparated tryptic digests of three glycoproteins taken as model compounds. Even glycopeptides containing short peptide backbones and large carbohydrate moieties gave high signal intensities when using the ILM though they did not appear in the THAP spectra. In the second set of experiments, the total tryptic digests were treated with endoglycosidase PNGase F to cleave off the N-linked glycans. When using the GTHAP matrix, it was possible to detect the glycans with high intensities in the presence of the tryptic peptides, whereas glycan ionization was completely suppressed when measured with the solid matrix THAP. The extent of metastable decay of glycopeptides was reduced when using the ILM. Altogether, GTHAP proved as a useful ILM particularly being superior to solid matrices in the context of glycosylation analysis.
Ullmer
Roman
R
Institute of Analytical Chemistry and Food Chemistry, University of Vienna, Währinger Strasse 38, A-1090 Vienna, Austria.
Rizzi
Andreas M
AM
D000098:Acetophenones / Q000737:chemistry; D000818:Animals; D000991:Antithrombins / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D006020:Glycopeptides / Q000737:chemistry* / Q000378:metabolism; D006146:Guanidines / Q000737:chemistry; D006801:Humans; D052578:Ionic Liquids / Q000737:chemistry*; D009961:Orosomucoid / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism*; D000509:alpha-Fetoproteins / Q000737:chemistry / Q000378:metabolism
D000098:Acetophenones; D000991:Antithrombins; D006020:Glycopeptides; D006146:Guanidines; D052578:Ionic Liquids; D009961:Orosomucoid; D010446:Peptide Fragments; D011134:Polysaccharides; D000509:alpha-Fetoproteins; C107067:2,4,6-trihydroxyacetophenone; D014357:Trypsin; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; C477069:1,1,3,3-tetramethylguanidine