How can temperature affect the stability of GFP during purification?

The correct answer highlights a critical aspect of protein chemistry and the behavior of GFP under varying temperature conditions. Green Fluorescent Protein (GFP) is a sensitive macromolecule that can undergo structural changes when exposed to heat. High temperatures may disrupt the hydrogen bonds, hydrophobic interactions, and other stabilizing forces that maintain the protein's three-dimensional structure. This disruption can lead to denaturation, causing the protein to unfold and lose its functional conformation, which directly affects its ability to fluoresce.

When proteins such as GFP denature, they not only lose their structural integrity but also their functional properties, including fluorescence. This process diminishes the protein's activity and can complicate its purification. Therefore, control of temperature is crucial during the purification process to ensure that the protein remains stable and retains its fluorescence properties, making it easier to isolate and study.