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Proteomics strategy based on liquid‐phase IEF and 2‐D DIGE: Application to bone marrow mesenchymal progenitor cells
Author(s) -
Seshi Beerelli
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600868
Subject(s) - proteomics , progenitor cell , mesenchymal stem cell , bone marrow , microbiology and biotechnology , biology , chemistry , stem cell , biochemistry , immunology , gene
Global comparative proteomics is a promising new approach with broad application in basic and clinical biological science. Recent advances include the development of 2‐D DIGE, a proteomic equivalent to mRNA differential display, in which differentially labeled samples are multiplexed and analyzed by high‐resolution 2‐DE. This study presents a new 2‐D DIGE protocol, in which complex protein samples from normal and leukemic human bone marrow mesenchymal progenitor cells were used as model samples for a novel combination of liquid‐phase IEF with 2‐D DIGE. Using liquid‐phase IEF, the normal and leukemic cells were pre‐fractionated into five subproteomes after multiplexing but prior to DIGE. Under these conditions, 2‐D DIGE resolved >5000 protein‐containing spots within the pH range 4.6–7.0. Differential labeling combined with subsequent MALDI‐MS/MS identified proteins that were differentially expressed in leukemic cells. This analysis mapped protein identities to 128 mesenchymal progenitor cell proteins with at least one unique peptide match at >95% confidence. Of these proteins, 72 (56%) were expressed more than 1.25‐fold higher or lower in leukemic cells compared with normal cells ( p <0.05). These data were used to infer gene ontology biological processes that may be altered in leukemic bone marrow mesenchymal progenitor cells.