While perusing the scientific literature late one night, youcome across a report that a protein called Thingamajig is thoughtto be involved in the development of antibiotic resistance, anemerging global health crisis. This putative involvement appears toinvolve an interaction with another protein, Whosiewhatsit. Youwould like to study Thingamajig, and its interaction withWhosiewhatsit, but neither protein has been cloned.
a) How would you set out to study these proteins and theirinteraction? Describe in general terms (a flowchart, for example),filling in the details once this framework is assembled. To get youstarted, how would you obtain the information necessary to expressand purify these proteins, and how would you manipulate thisinformation?
b) Assuming Thingamajig is positively charged and Whosiewhatsitis negatively charged, what purification approach might you use topurify each of them independently? What approach might you take topurify the complex of the two?
c) What approach would you take to identify the amino acidresidues — in both Thingamajig and Whosiewhatsit — that underlietheir interaction with one another?
