The Mechanism of 2,2,2-Trichloroacetic Acid-Induced Protein Precipitation |
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Authors: | T Sivaraman T K S Kumar G Jayaraman and C Yu |
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Institution: | (1) Department of Chemistry, National Tsing Hua University, Hsinchu, Taiwan |
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Abstract: | The mechanism of 2,2,2-trichloroacetic acid (TCA)-induced precipitation of proteins is studied. The TCA-induced protein precipitation curves are observed to be U-shaped. It is bound that the protein-precipitate-inducing effects of TCA are due to the three chloro groups in the molecule. Using cardiotoxin III (CTX III) isolated from the Taiwan cobra (Naja naja atra), as a model protein, we attempt to understand the molecular basis for the TCA-induced effects. Employing circular dichroism, proton–deuterium exchange in conjunction with conventional 2D NMR techniques, and 1-anilino naphthalene-8-sulfonate-binding experiments, we demonstrate that CTX III is in a partially structured state similar to the A state in 3% w/v TCA. It is postulated that the formation of this sticky partial structured A state in the TCA-induced unfolding pathway is responsible for the acid-induced protein precipitation. |
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Keywords: | A-state CTX III protein precipitation TCA |
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