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Determining the subunit composition of a protein from data

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You isolate a new protein. Column chromatography, in Tris-HCl buffer pH 7.4, demonstrates that this protein has a molecular weight of 110,000 D. Chromatography in the presence of 6 M guanidine hydrochloride demonstrates two peaks with masses of approximately 13,000 and approximately 26,000 D. The peak at approximately 26,000 D contains 80.3%, by mass, of the protein. The protein from this peak, when analyzed using isoelectric focusing column chromatography demonstrates that this peak contains two different proteins, each being 50% of the mass of this peak and both with acidic pIâ??s. These two 26,000 D proteins are separately treated with a^-mercaptoethanol and analyzed by column chromatography in the presence of 6 M guanidine hydrochloride. This demonstrates equal amounts of three peptides with molecular weight of 10,000 D, 9,000 D, and 7,500 D for one protein and 17,500 and 9,000 for the other protein. From these data, determine what the composition of this new protein is.

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Solution Summary

In this solution we explore how to elucidate the number and types of subunits of a multi-unit protein from biochemical data. A detailed approach to the problem is provided in the solution.

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This should be a multi-subunit protein with the following composition:

2 of the 13 kDa subunits (since the first two peaks showed an approximate 80% to 20%, the 20% being the 13000 D weight. 80% was the 26 kDa peak).

The 26 kDa protein is further broken up ...

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