Application of On-Line nanoLC-IT-TOF in the Identification of Serum beta-Catenin Complex in Mice Scald Model

• 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.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Medicinsk genetik (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Medical Genetics (hsv//eng)

Nyckelord

Dependent Protein-Kinase

Cancer

Proliferation

Activation

Pathway

Growth

Phosphorylation

Stabilization

Inhibition

Carcinoma

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