Determination of the Abundance Proteins of Adipose Synthesis and β-Oxidation Using ¹⁸O Isotope Labeled and QconCAT Protein Combined with Multiple Reaction Monitoring Mass Spectrometry

A novel method using QconCAT protein combined with ¹⁸O-labeling technique and multiple reaction monitoring mass spectrometry was established to quantify proteins involved in the pathway of adipose synthesis and β-oxidation. Firstly, the QconCAT proteins were constructed, and expressed in Escherichia coli. The digestion and the ¹⁸O labeling efficiency were optimized. Secondly, the novel method of quantifying adipose synthesis and β-oxidation was developed. Lastly, target proteins of obesity and control groups were quantified by MRM. The results showed that the abundance of the enoyl ACP reductase and malonyl-COA ACP transferase increased significantly in the obesity group with P<0.05. It is consistent to previous reports and means the method had its reliability to analyze the biological sample.