Application of On-Line nanoLC-IT-TOF in the Identification of Serum beta-Catenin Complex in Mice Scald Model
2012 (English)In: PLoS ONE, ISSN 1932-6203, Vol. 7, no 10, e46530- p.Article in journal (Refereed) Published
Severe burn shock remains an unresolved clinical problem with an urgent need to explore novel therapeutic treatments. Intracellular beta-catenin, through interaction with other proteins, has been reported to be able to regulate the size of cutaneous wounds. Higher expression of beta-catenin is associated with larger sized wounds. However, the identification of serum beta-catenin complex is difficult and has been rarely reported. The exploitation of more binding partners can contribute to uncovering the exact mechanisms behind serum beta-catenin mediated biological effects. Here, we describe a method that consists of immunoprecipitation, SDS-PAGE, in-gel digestion, and nanoLC coupled to LCMS-IT-TOF for the investigation of serum beta-catenin complex in mice scald model. Among selected gel bands obtained from the protein gels, a total of 31 peptides were identified and sequenced with high statistical significance (p<0.01). Three proteins (alpha-2-marcoglobulin, serine protease inhibitor A3K, and serine protease inhibitor A1A) were identified and validated with high reliability and high reproducibility. It was inferred that these proteins might interact with serum beta-catenin, which could affect the wound healing resulting from burn shock. Our study demonstrated that the on-line coupling of nano-LC with a LCMS-IT-TOF mass spectrometer was capable of sensitive and automated characterization of the serum beta-catenin complex in mice scald model.
Place, publisher, year, edition, pages
2012. Vol. 7, no 10, e46530- p.
Dependent Protein-Kinase, Cancer, Proliferation, Activation, Pathway, Growth, Phosphorylation, Stabilization, Inhibition, Carcinoma
IdentifiersURN: urn:nbn:se:kth:diva-105002DOI: 10.1371/journal.pone.0046530ISI: 000309889400014ScopusID: 2-s2.0-84867294607OAI: oai:DiVA.org:kth-105002DiVA: diva2:570076
QC 201211162012-11-162012-11-152012-11-16Bibliographically approved