To analyze the denaturing effect of urea, we plotted absorbance vs. concentratio

Question

To analyze the denaturing effect of urea, we plotted absorbance vs. concentration in the protein folding lab. Use data table and plot below to calculate the ratio of fD to fN (for fD fration denatured protein and fN fraction of protein are in native form). Calculate the fD to fN ratio at urea concentrations 5.25, 6, 6.75 (all values in M). Also, estimate urea concentration at transition midpoint when fD=fN.

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[urea] 0 .75 1.5 2.25 3 3.75 4.5 5.25 6 6.75 7.5 Absorbance 1.233 1.239 1.22 1.269 1.2 1.229 1.198 1.151 .921 .496 .350 To analyze the denaturing effect of urea, we plotted absorbance vs. concentration in the protein folding lab. Use data table and plot below to calculate the ratio of fD to fN (for fD fration denatured protein and fN fraction of protein are in native form). Calculate the fD to fN ratio at urea concentrations 5.25, 6, 6.75 (all values in M). Also, estimate urea concentration at transition midpoint when fD=fN.

Explanation / Answer

Some assumptions:

-the native protein is responsible for absorption

-in the first experiment (without urea) the entire protein is in native form

-the denaturated protein is in some way not absorbing

-the calibration curve is linear, so one can write:

Absorbance A = k (a constant)

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