期刊
JOURNAL OF PROTEOME RESEARCH
卷 14, 期 2, 页码 654-663出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr501076j
关键词
proteomics; peptidomics; Alzheimer's disease; neurodegenerative disease; biomarker discovery; cerebrospinal fluid; isobaric labeling; quantification; clinical protemics
资金
- Swedish Research Council
- Knut and Alice Wallenberg Foundation
- Vetenskapsradet [521-2011-4709]
- EMIF-AD
- Emil och Wera Cornells stiftelse
- Aina Wallstrom och Mary-Ann Sjobloms stiftelse
- Demensforbundet
- Magnus Bergvalls stiftelse
- Adlerbertska stiftelsen
- Stiftelsen for Gamla Tjanarinnor
- Gun och Bertil Stohnes stiftelse
- Kungl och Hvitfeldtska stiftelsen
- Wilhelm och Martina Lundgrens vetenskapsfond
- Emil and Maria Palm Foundation
- Stiftelsen Greta Johansson och Brita Anderssons minnesfond
Many disease processes in the brain are reflected in the protein composition of the cerebrospinal fluid (CSF). In addition to proteins, CSF also contains a large number of endogenous peptides whose potential as disease biomarkers largely remains to be explored. We have developed a novel workflow in which multiplex isobaric labeling is used for simultaneous quantification of endogenous CSF peptides and proteins by liquid chromatography coupled with mass spectrometry. After the labeling of CSF samples, endogenous peptides are separated from proteins by ultrafiltration. The proteins retained on the filters are trypsinized, and the tryptic peptides are collected separately. We evaluated this technique in a comparative pilot study of CSF peptide and protein profiles in eight patients with Alzheimer's disease (AD) and eight nondemented controls. We identified several differences between the AD and control group among endogenous peptides derived from proteins known to be associated with AD, including neurosecretory protein VGF (ratios AD/controls 0.45-0.81), integral membrane protein 2B (ratios AD/controls 0.72-0.84), and metallothionein-3 (ratios AD/controls 0.51-0.61). Analysis of tryptic peptides identified several proteins that were altered in the AD group, some of which have previously been reported as changed in AD, for example, VGF (ratio AD/controls 0.70).
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