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Exploring the CSF Phosphoproteome with a Novel Fluorescent In‐Gel Stain
Author(s) -
Loyal Brian,
Stone David,
Turner Jeff,
Boland Judy,
Crawford Angela,
Chen Dian Er
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a632-e
Subject(s) - cerebrospinal fluid , stain , cerebral spinal fluid , chemistry , gel electrophoresis , microbiology and biotechnology , staining , biology , computational biology , chromatography , pathology , neuroscience , medicine , biomedical engineering
Human Cerebrospinal Fluid (CSF) is a serum‐like solution that bathes the brain and spinal column and provides an important source of potential disease biomarkers. Phosphorylated CSF proteins may supply information about a variety of neuronal pathologies, including Alzheimer's disease and paraneoplastic cerebellar degeneration. Current methods of identifying these phosphorylated biomarkers rely on immunoassays and require specific prior knowledge about the target protein. One alternative is the use of a global indicator for phosphoproteins, such as the PhosDecor ™ fluorescent stain. This allows the potential visualization of an entire CSF phosphoproteome using either 2‐dimensional electrophoresis (2DE) or another gel‐based system. In this study, we utilized the PhosDecor stain to identify phosphorylated proteins in‐gel from samples of (a) whole CSF, and (b) CSF processed with the ProteoPrep® 20 immunodepletion technology. In the latter case, several abundant protein species were removed from the sample, allowing increased visualization of scarce proteins of interest. After separation by 2DE, relevant phosphoproteins were excised for mass spectrometric analysis and identification. As a result, we report on a novel survey of the detailed CSF phosphoproteome.