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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Morschheuser, Lena Mükusch, Sandra Riedner, Maria Seitz, Harald Rohn, Sascha |
| Description | Author Affiliation: Morschheuser L ( University of Hamburg, Hamburg School of Food Science, Institute of Food Chemistry, Grindelallee 117, D-20146 Hamburg, Germany. Electronic address: lena.morschheuser@chemie.uni-hamburg.de.); Mükusch S ( Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytic and Bioprocesses, Am Mühlenberg 13, D-14476 Potsdam/Golm, Germany); Riedner M ( University of Hamburg, Institute of Organic Chemistry, Martin-Luther-King-Platz 6, D-20146 Hamburg, Germany. Electronic address: maria.riedner@chemie.uni-hamburg.de.); Seitz H ( Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytic and Bioprocesses, Am Mühlenberg 13, D-14476 Potsdam/Golm, Germany. Electronic address: harald.seitz@izi-bb.fraunhofer.de.); Rohn S ( University of Hamburg, Hamburg School of Food Science, Institute of Food Chemistry, Grindelallee 117, D-20146 Hamburg, Germany. Electronic address: rohn@chemie.uni-hamburg.de.) |
| Abstract | This study aimed at developing a rapid chromatographic assay to monitor phosphorylation sites in peptides. For the analysis of nociceptive signal transduction pathways, the detection of phosphorylated proteins/peptides plays a fundamental role. To get further insights in the phosphorylation mechanism of protein kinase C-ε (PKC-ε) and protein kinase A (PKA), potential targets were divided into subsections resulting in peptides that contain only one possible phospho-binding site. The use of high-performance thin-layer chromatography (HPTLC) offers the possibility of a high throughput of samples and the advantage of a quick sample clean-up. A combined strategy of an effect-directed overlay procedure on the TLC plate using specific antibodies (immunostaining, HPTLC-IS) as well as a parallel, direct mass spectrometric methodology by HPTLC-MALDI-TOF-MS was developed. With regard to HPTLC-IS, validation of the data exhibited a lower limit of detection than the traditionally used protein derivatization reagent fluorescamine. Besides the identification of the phosphorylated peptides, a semi-quantitative estimation can be performed with HPTLC-IS. |
| File Format | HTM / HTML |
| ISSN | 15700232 |
| Volume Number | 1008 |
| e-ISSN | 1873376X |
| Journal | Journal of Chromatography B |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-01-01 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Chemistry Chromatography, Thin Layer Methods Peptides Analysis Amino Acid Sequence Molecular Sequence Data Chemistry Phosphorylation Spectrometry, Mass, Matrix-assisted Laser Desorption-ionization Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry |
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