Dataset Information

Proteome analysis of the MO3.13 cell line from human oligodendrocytes

ABSTRACT: MO3.13 is an immortal human-human hybrid cell line that express phenotypic characteristics of primary oligodendrocytes. It was created by fusing a 6-thioguanine-resistant mutant of the human rhabdomyosarcoma RD (cancer of skeletal muscle) with adult human oligodendrocytes by a lectin-enhanced polyethylene glycol procedure. In contrast to the tumor parent, MO3.13 expressed surface immunoreactivity for galactosyl cerebroside(GS) and intracellular immunoreactivity for myelin basic protein (MBP), proteolipid protein (PLP), and glial fibrillary acidic protein (GFAP). MO3.13 also exhibit the markers of immature oligodendrocytes GalC (galactosylceramidase) and CNPase. Upon differentiation, the MO3.13 cells have been also shown to express the MBP and MOG markers. Data analysis: Each MS raw file was processed using ProteoWizard for the generation of a MGF file. These were processed in SearchGUI, which runs the search engines Open Mass Spectrometry Search Algorithm (OMSSA) and X!Tandem against the UniProt human protein database (release 2013_08, 20,266 sequences). Â Search parameters were: peptide and fragment ion mass accuracy 10 ppm and 0.5 Da, respectively; protein and peptide FDRs 1%; two miss cleavages; trypsin as enzyme; fixed modifications: cysteine carbamidomethylation; variable modifications: methionine oxidation. In order to generate one single proteome dataset of MO3.13, resulting data was processed in PeptideShaker.

INSTRUMENT(S): LTQ Orbitrap, MS:1000031

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: None Listed

PROVIDER: MSV000083971 | MassIVE | Wed Jun 12 21:27:00 BST 2019

SECONDARY ACCESSION(S): PXD000263

REPOSITORIES: MassIVE

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The human oligodendrocyte proteome.

Iwata Keiko K Café-Mendes Cecilia C CC Schmitt Andrea A Steiner Johann J Manabe Takayuki T Matsuzaki Hideo H Falkai Peter P Turck Christoph W CW Martins-de-Souza Daniel D

Proteomics 20131120 23-24

Myelination of the CNS is performed by oligodendrocytes (OLs), which have been implicated in brain disorders, such as multiple sclerosis and schizophrenia. We have used the human oligodendroglial cell line MO3.13 to establish an OL reference proteome database. Proteins were prefractionationated by SDS-PAGE and after in-gel digestion subjected to nanoflow LC-MS analysis. Approximately 11 600 unique peptides were identified and, after stringent filtering, resulted in 2290 proteins representing nin ...[more]

PMID: 24167090

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