You have recently begun studying pulmonary disease in a lab that specializes in

Question

You have recently begun studying pulmonary disease in a lab that specializes in culturing human lung epithelial cells. Specifically, you are interested in how the protease trypsin affects the connections that form between lung epithelial cells. Your advisor instructs you to purify an 8400 dalton trypsin inhibitor known as the Bowman-Birk inhibitor (BBI) from soybean meal. Following mechanical cell lysis and centrifugation, you perform a series of column chromatography steps to purify BBI from the soluble fraction. First, you perform anion-exchange chromatography followed by analysis of collected fragments using spectrophotometry (where absorbance of 280 light indicates the presence of protein). Are the proteins in fraction 45 more or less negative than proteins found in fraction 54? Explain your answer

Explanation / Answer

The proteins in 45 is more negative than those in 54. Because absorbance at 280nm is highest for tryptophan which is slightly more negative and hence the charge of the protein is more negative.

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