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The ribonuclease A protein has 8 Cys residues and makes 4 disulfide bo.pdf

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The ribonuclease A protein has 8 Cys residues and makes 4 disulfide bonds in the native folded conformation. If the protein is allowed to make disulfide bonds while it is in the denatured (completely unfolded) state, the number of possible configurations with 4 disulfide pairings is found to be 105 . (a) Rationalize the 105 configurations. (b) The protein is unfolded by addition of urea and the disulfide bonds are disrupted by addition of a reducing agent. The protein is then allowed to fold back by (i) first adding an oxidizing agent (to reform disulfide bonds) before removing urea (ii) first removing urea and then adding the oxidizing agent In each of the scenarios above, what level of protein activity is recovered? Explain..

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The ribonuclease A protein has 8 Cys residues and makes 4 disulfide bonds in the native folded conformation. If the protein is allowed to make disulfide bonds while it is in the denatured (completely unfolded) state, the number of possible configurations with 4 disulfide pairings is found to be 105 . (a) Rationalize the 105 configurations. (b) The protein is unfolded by addition of urea and the disulfide bonds are disrupted by addition of a reducing agent. The protein is then allowed to fold back by (i) first adding an oxidizing agent (to reform disulfide bonds) before removing urea (ii) first removing urea and then adding the oxidizing agent In each of the scenarios above, what level of protein activity is recovered? Explain.

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The ribonuclease A protein has 8 Cys residues and makes 4 disulfide bo.pdf

  • 1. The ribonuclease A protein has 8 Cys residues and makes 4 disulfide bonds in the native folded conformation. If the protein is allowed to make disulfide bonds while it is in the denatured (completely unfolded) state, the number of possible configurations with 4 disulfide pairings is found to be 105 . (a) Rationalize the 105 configurations. (b) The protein is unfolded by addition of urea and the disulfide bonds are disrupted by addition of a reducing agent. The protein is then allowed to fold back by (i) first adding an oxidizing agent (to reform disulfide bonds) before removing urea (ii) first removing urea and then adding the oxidizing agent In each of the scenarios above, what level of protein activity is recovered? Explain.