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    High-throughput proteomics detection of novel splice isoforms in human platelets.

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    Authors
    Power, Karen A
    McRedmond, James P
    de Stefani, Andreas
    Gallagher, William M
    Gaora, Peadar O
    Affiliation
    UCD Conway Institute and UCD School of Biomolecular & Biomedical Sciences, UCD Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
    Issue Date
    2009
    MeSH
    Amino Acid Sequence
    Blood Platelets
    Databases, Factual
    Exons
    Genome, Human
    Humans
    Mass Spectrometry
    Metalloendopeptidases
    Peptides
    Protein Isoforms
    Proteomics
    RNA, Messenger
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    Citation
    High-throughput proteomics detection of novel splice isoforms in human platelets. 2009, 4 (3):e5001 PLoS ONE
    Journal
    PloS one
    URI
    http://hdl.handle.net/10147/94732
    DOI
    10.1371/journal.pone.0005001
    PubMed ID
    19308253
    Abstract
    Alternative splicing (AS) is an intrinsic regulatory mechanism of all metazoans. Recent findings suggest that 100% of multiexonic human genes give rise to splice isoforms. AS can be specific to tissue type, environment or developmentally regulated. Splice variants have also been implicated in various diseases including cancer. Detection of these variants will enhance our understanding of the complexity of the human genome and provide disease-specific and prognostic biomarkers. We adopted a proteomics approach to identify exon skip events - the most common form of AS. We constructed a database harboring the peptide sequences derived from all hypothetical exon skip junctions in the human genome. Searching tandem mass spectrometry (MS/MS) data against the database allows the detection of exon skip events, directly at the protein level. Here we describe the application of this approach to human platelets, including the mRNA-based verification of novel splice isoforms of ITGA2, NPEPPS and FH. This methodology is applicable to all new or existing MS/MS datasets.
    Language
    en
    ISSN
    1932-6203
    ae974a485f413a2113503eed53cd6c53
    10.1371/journal.pone.0005001
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