200401M2-2 - Chem 237 Midterm#2 from 1 Isolated-subunits of...

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Chem 237 Midterm #2 from March 10, 2004 1. Isolated β -subunits of HbA (normal hemoglobin) can be reassembled into β 4 tetramers using a specific set of conditions (you do not need to know these conditions to do the problem). These β 4 tetramers have a P 50 value of 6 torr and show no cooperativity. The isolated α - subunits of HbA have a pI of 7.4, and the β -subunits have a pI of 6.9. a. Assuming that you have purified HbA and have a method to selectively break the quaternary interactions between the α and β subunits, how might you separate the β - subunits from the α -subunits, using one of the separation techniques we have talked about in class? Be sure to include any pertinent details about the method you use (resin type, pH, salt concentration, etc). b. Draw an O 2 binding curve for the β 4 tetramers; be sure to label the axes properly, and include an Mb and Hb curve for reference. Why did you draw the curve the way you did?
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