Figure 7.
Structural modeling to gain insights into the engineered mutations of PHL7
(A) AlphaFold predicted structures of PHL7-Jemez (magenta) aligned to the crystal structure of PHL7-WT (green). The regions of maximum variability in the backbone were observed in the loops consisting of residues 46–54 and 111–124. Published sequence PHL7-L93F/Q95Y, was also modeled and overlayed on the crystal structure of PHL7-WT. The catalytic triad residues S131, D177, and H209 are highlighted in cyan.
(B) ROSETTA modeling shows stabilizing effect of Q→E and R→K mutations at positions 175 and 205, respectively. A hydrogen bond is formed only in the case of the double mutation Q175E and R205K in PHL7-Santa Fe. The side chains of residues 175 and 205 are shown for PHL7-WT (left), PHL7-Jemez (middle), and PHL7-Santa Fe (right).
