title: Top-down label-free LC-MALDI analysis of the peptidome during neural progenitor cell differentiation reveals complexity in cytoskeletal protein dynamics and identifies progenitor cell markers.
abstract: In the field of stem cell research, there is a strong requirement for the discovery of new biomarkers that more accurately define stem and progenitor cell populations, as well as their differentiated derivatives. The very-low-molecular-weight (<5 kDa) proteome/peptidome remains a poorly investigated but potentially rich source of cellular biomarkers. Here we describe a label-free LC-MALDI-TOF/TOF quantification approach to screen the very-low-molecular-weight proteome, i.e. the peptidome, of neural progenitor cells and derivative populations to identify potential neural stem/progenitor cell biomarkers. Twelve different proteins were identified on the basis of MS/MS analysis of peptides, which displayed differential abundance between undifferentiated and differentiated cultures. These proteins included major cytoskeletal components such as nestin, vimentin, and glial fibrillary acidic protein, which are all associated with neural development. Other cytoskeletal proteins identified were dihydropyrimidinase-related protein 2, prothymosin (thymosin α-1), and thymosin β-10. These findings highlight novel stem cell/progenitor cell marker candidates and demonstrate proteomic complexity, which underlies the limitations of major intermediate filament proteins long established as neural markers.
authors.affiliation: School of Biological and Biomedical Sciences, Durham University, Durham, UK.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Maltman
authors.forename: Sven
authors.initials: S
authors.lastname: Brand
authors.forename: Eckhard
authors.initials: E
authors.lastname: Belau
authors.forename: Rainer
authors.initials: R
authors.lastname: Paape
authors.forename: Detlev
authors.initials: D
authors.lastname: Suckau
authors.forename: Stefan A
authors.initials: SA
authors.lastname: Przyborski
mesh_terms: D000595:Amino Acid Sequence; D015415:Biomarkers; D015153:Blotting, Western; D002454:Cell Differentiation; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D055495:Neurogenesis; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology*
chemical_list: D015415:Biomarkers; D003598:Cytoskeletal Proteins