![]() In our opinion, the findings of this study provide useful clues toward development of efficient protocols for the isolation and analysis of the entire proteome. The results of the study clearly demonstrate that TCA-induced protein precipitation occurs due to the reversible association of the “MG-like” partially structured intermediate state(s). 1H- 15N chemical shift perturbation data obtained using NMR spectroscopy indicate that interactions stabilizing the β-strands at the N- and C- terminal ends (of aFGF) are disrupted in the trichloroacetate-induced “MG-like” state. ![]() Urea-induced unfolding and limited proteolytic digestion data reveal that the partially structured intermediate is significantly less stable than the native conformation. Results of the 1-anilino-8-napthalene sulfonate (ANS) and size-exclusion chromatography, obtained using acidic fibroblast growth factor (aFGF), show that a stable “molten globule-like” partially structured intermediate accumulates maximally in 5% (w/v) of trichloroacetate. TCA is significantly less effective in precipitating unfolded states of proteins. TCA-induced protein precipitation curves are U-shaped and the shape of the curve is observed to be independent of the physicochemical properties of proteins. ![]() In this study, we examine the mechanism of the TCA-induced protein precipitation reaction. Sample preparation for proteomic analysis involves precipitation of protein using 2,2,2-trichloroacetic acid (TCA). ![]()
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