Determination of Phosphorus-, Copper-, and Zinc-Containing Human Brain Proteins by LA-ICPMS and MALDI-FTICR-MS
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- 作者：J. Sabine Becker ; Miroslav Zoriy ; J. Susanne Becker ; Carola Pickhardt ; Eugen Damoc ; Gabor Juhacz ; Miklos Palkovits ; and Michael Przybylski
- 刊名：Analytical Chemistry
- 出版年：2005
- 出版时间：September 15, 2005
- 年：2005
- 卷：77
- 期：18
- 页码：5851 - 5860
- 全文大小：539K
- 年卷期：v.77,no.18(September 15, 2005)
- ISSN：1520-6882
文摘
Human brain proteins containing phosphorus, copper,and zinc were detected directly in protein spots in gels ofa human brain sample after separation by two-dimensional gel electrophoresis using laser ablation inductivelycoupled plasma mass spectrometry (LA-ICPMS). A powerful laser ablation system with cooled laser ablationchamber was coupled to a double-focusing sector fieldICPMS. The separated protein spots in 2D gels were fastscreened using the optimized microanalytical LA-ICPMStechnique measured at medium mass resolution with afocused laser beam (wavelength, 213 nm; diameter oflaser crater, 50 
m; and laser power density, 3 × 109 Wcm-2) with respect to selected three essential elements.Of 176 protein spots in 2D gel from a human brainsample, phosphorus, copper, and zinc were detected in31, 43, and 49 protein spots, respectively. For the firsttime, uranium as a naturally occurring radioactive element was found in 20 selected protein spots. The detection limits for P, S, Cu, Zn and U were determined insingular protein spots with 0.0013, 1.29, 0.029, 0.063,and 0.000 01 mg g-1, respectively. A combination of LA-ICPMS with matrix-assisted laser desorption/ionizationFourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR-MS) was applied for the identification of selected protein spots from human brainprotein separated by 2D gel electrophoresis. CombiningMALDI-FTICR-MS for the structure analysis of metal- andphosphorus-containing human brain proteins with LA-ICPMS, the direct analysis of heteroelements on separated proteins in 2D gels can be performed. For quantification of analytical LA-ICPMS data, the number of sulfuratoms per protein (and following the sulfur concentration)determined by MALDI-FTICR-MS was used for internalstandardization. From the known sulfur concentration inprotein, the concentration of other heteroelements wascalculated. In addition, the number of phosphorylationand the phosphorylation sites of phosphorylated proteinsin the human brain sample detected by LA-ICPMS weredetermined by MALDI-FTICR-MS. This technique allowsthe study of posttranslational modifications in humanbrain proteins.
m; and laser power density, 3 × 109 Wcm-2) with respect to selected three essential elements.Of 176 protein spots in 2D gel from a human brainsample, phosphorus, copper, and zinc were detected in31, 43, and 49 protein spots, respectively. For the firsttime, uranium as a naturally occurring radioactive element was found in 20 selected protein spots. The detection limits for P, S, Cu, Zn and U were determined insingular protein spots with 0.0013, 1.29, 0.029, 0.063,and 0.000 01 mg g-1, respectively. A combination of LA-ICPMS with matrix-assisted laser desorption/ionizationFourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR-MS) was applied for the identification of selected protein spots from human brainprotein separated by 2D gel electrophoresis. CombiningMALDI-FTICR-MS for the structure analysis of metal- andphosphorus-containing human brain proteins with LA-ICPMS, the direct analysis of heteroelements on separated proteins in 2D gels can be performed. For quantification of analytical LA-ICPMS data, the number of sulfuratoms per protein (and following the sulfur concentration)determined by MALDI-FTICR-MS was used for internalstandardization. From the known sulfur concentration inprotein, the concentration of other heteroelements wascalculated. In addition, the number of phosphorylationand the phosphorylation sites of phosphorylated proteinsin the human brain sample detected by LA-ICPMS weredetermined by MALDI-FTICR-MS. This technique allowsthe study of posttranslational modifications in humanbrain proteins.