Phosphopeptide screening using nanocrystalline titanium dioxide films as affinity matrix-assisted laser desorption ionization targets in mass spectrometry
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Phosphopeptide screening using nanocrystalline titanium dioxide films as affinity matrix-assisted laser desorption ionization targets in mass spectrometry. / Niklew, Marie-Luise; Hochkirch, Ulrike; Melikyan, Anna; Moritz, Thomas; Kurzawski, Sandra; Schlüter, Hartmut; Ebner, Ingo; Linscheid, Michael W.
in: ANAL CHEM, Jahrgang 82, Nr. 3, 3, 01.02.2010, S. 1047-1053.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Phosphopeptide screening using nanocrystalline titanium dioxide films as affinity matrix-assisted laser desorption ionization targets in mass spectrometry
AU - Niklew, Marie-Luise
AU - Hochkirch, Ulrike
AU - Melikyan, Anna
AU - Moritz, Thomas
AU - Kurzawski, Sandra
AU - Schlüter, Hartmut
AU - Ebner, Ingo
AU - Linscheid, Michael W
PY - 2010/2/1
Y1 - 2010/2/1
N2 - The use of nanocrystalline titanium dioxide films as affinity targets for the selective isolation and enrichment of phosphopeptides with subsequent analysis by matrix-assisted laser desorption ionization (MALDI) mass spectrometry is described. A strong affinity of phosphopeptides to anatase titanium dioxide surfaces is observed, and a standard protocol for the selective isolation and enrichment of phosphopeptides on titanium dioxide films using a proteolytic digest of alpha- and beta-casein was developed. All washing and elution procedures using these films can be processed directly on the MALDI target, thereby avoiding sample contamination and losses. In addition, the enrichment of the phosphopeptides was improved due to a considerable enlargement of the surface. Several film substrates compatible with routine inlet systems of mass spectrometers, as conductive glass, aluminum, and silicon, have been manufactured and tested. A biological application was examined by the human fibrinogen-thrombin system. For a quantification and comparison of different expression levels of phosphoproteins in biological systems, the peptides were labeled with S-methyl thioimidate reagents. The capability of this method for high-throughput applications make the use of mesoporous titanium dioxide films as an affinity MALDI target a promising tool in phosphoproteomics. A combination of an amidation protocol showed that a quantification of phosphorylated peptides can easily be performed using TiO(2) films.
AB - The use of nanocrystalline titanium dioxide films as affinity targets for the selective isolation and enrichment of phosphopeptides with subsequent analysis by matrix-assisted laser desorption ionization (MALDI) mass spectrometry is described. A strong affinity of phosphopeptides to anatase titanium dioxide surfaces is observed, and a standard protocol for the selective isolation and enrichment of phosphopeptides on titanium dioxide films using a proteolytic digest of alpha- and beta-casein was developed. All washing and elution procedures using these films can be processed directly on the MALDI target, thereby avoiding sample contamination and losses. In addition, the enrichment of the phosphopeptides was improved due to a considerable enlargement of the surface. Several film substrates compatible with routine inlet systems of mass spectrometers, as conductive glass, aluminum, and silicon, have been manufactured and tested. A biological application was examined by the human fibrinogen-thrombin system. For a quantification and comparison of different expression levels of phosphoproteins in biological systems, the peptides were labeled with S-methyl thioimidate reagents. The capability of this method for high-throughput applications make the use of mesoporous titanium dioxide films as an affinity MALDI target a promising tool in phosphoproteomics. A combination of an amidation protocol showed that a quantification of phosphorylated peptides can easily be performed using TiO(2) films.
KW - Animals
KW - Humans
KW - Cattle
KW - Amino Acid Sequence
KW - Molecular Sequence Data
KW - Nanoparticles/chemistry
KW - Trypsin/metabolism
KW - Caseins/chemistry/metabolism
KW - Fibrinogen/chemistry/metabolism
KW - Phosphopeptides/analysis
KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
KW - Thrombin/metabolism
KW - Titanium/chemistry
KW - Animals
KW - Humans
KW - Cattle
KW - Amino Acid Sequence
KW - Molecular Sequence Data
KW - Nanoparticles/chemistry
KW - Trypsin/metabolism
KW - Caseins/chemistry/metabolism
KW - Fibrinogen/chemistry/metabolism
KW - Phosphopeptides/analysis
KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
KW - Thrombin/metabolism
KW - Titanium/chemistry
U2 - 10.1021/ac902403m
DO - 10.1021/ac902403m
M3 - SCORING: Journal article
C2 - 20067263
VL - 82
SP - 1047
EP - 1053
JO - ANAL CHEM
JF - ANAL CHEM
SN - 0003-2700
IS - 3
M1 - 3
ER -