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For the D101/Mg2+ functional unit (left), the black residues and Zn2+ ion represent a potential route for the energetic connections between these residues.
These studies include incisive double mutant cycle analysis demonstrating functional and energetic connections between residues (Hermes et al., 1990; Dion et al., 1993; Horovitz et al., 1994; Rajagopalan et al., 2002; Masterson et al., 2008; Singh et al., 2014).
In particular, there is no way to discern from structural inspection of an active site whether all neighbors have substantial energetic interactions, the scale and form of these energetic connections, or whether and how far beyond nearest neighbors functional effects extend.
Of particular interest will be understanding the interplay between the broader scaffold and the active site residues along with their functional roles and energetic connections, and such studies will ultimately rely on the development of methods that are high-throughput and highly quantitative.
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