Identification of alternatively spliced transcripts using a proteomic informatics approach.

We present the protocol for the identification of alternatively spliced peptide sequences from tandem mass spectrometry datasets searched using X!Tandem against our modified ECgene resource with all potential translation products and then matched with the Michigan Peptide to Protein Integration (MPPI) scheme. This approach is suitable for human and mouse datasets. Application of the method is illustrated with a study of the Kras activation-Ink4/Arf deletion mouse model of human pancreatic ductal adenocarcinoma.

[1]  L. Bracco,et al.  The relevance of alternative RNA splicing to pharmacogenomics. , 2003, Trends in biotechnology.

[2]  Rolf Apweiler,et al.  InterProScan: protein domains identifier , 2005, Nucleic Acids Res..

[3]  Patrick G. A. Pedrioli Trans-Proteomic Pipeline: A Pipeline for Proteomic Analysis , 2010, Proteome Bioinformatics.

[4]  T. Cooper,et al.  Pre-mRNA splicing and human disease. , 2003, Genes & development.

[5]  Namshin Kim,et al.  ECgene: genome-based EST clustering and gene modeling for alternative splicing. , 2005, Genome research.

[6]  R. Aebersold,et al.  Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry , 2001, Nature Biotechnology.

[7]  Damian Fermin,et al.  Identification of novel alternative splice isoforms of circulating proteins in a mouse model of human pancreatic cancer. , 2009, Cancer research.

[8]  J. Venables Aberrant and Alternative Splicing in Cancer , 2004, Cancer Research.

[9]  M. Garcia-Blanco,et al.  Alternative splicing in disease and therapy , 2004, Nature Biotechnology.

[10]  Thomas L. Madden,et al.  BLAST: at the core of a powerful and diverse set of sequence analysis tools , 2004, Nucleic Acids Res..

[11]  Damian Fermin,et al.  Novel gene and gene model detection using a whole genome open reading frame analysis in proteomics , 2006, Genome Biology.

[12]  Robertson Craig,et al.  TANDEM: matching proteins with tandem mass spectra. , 2004, Bioinformatics.

[13]  Jing Gao,et al.  Integrating and annotating the interactome using the MiMI plugin for cytoscape , 2009, Bioinform..

[14]  W. J. Kent,et al.  BLAT--the BLAST-like alignment tool. , 2002, Genome research.