FIGURE 4.

Comparison of predicted and experimental ΔΔG values indicates that none of a range of different computational approaches can accurately predict all tested mutations at C. thermocellum cohesin positions Leu⁸³, Asn³⁷, and Asp³⁹. The plot shows experimental and predicted ΔΔG values (y axis) of the different mutations (colored red for Leu⁸³, blue for Asp³⁹, and green for Asn³⁷). The x axis plots results from the experiment (exp) as well as from a range of different approaches, including different versions of Rosetta alanine scanning (Rosetta2.3 used in this study and a newly calibrated protocol Rosetta 3.0* (34)), FoldX (35), Hunter (37), two versions of the Orbit interface energy function EPPI-2 (36), and CC/PBSA (38). Incorrect predictions are boxed in bold black. All protocols accurately predict the effects of mutations of Leu⁸³ but perform less well on polar mutations. See “Experimental Procedures” for more details about the different protocols.