Structure and Function of the Refined C-Terminal Loop in Imidazole Glycerol Phosphate Dehydratase from Different Homologs.

IGPD is an essential metalloenzyme that catalyzes histidine biosynthesis. We found that its C-terminus loop region has a vital role in determining enzyme activity but has been hardly mentioned before. In this work, we focused on the dynamic feature and function of C-Loop in Arabidopsis thaliana and Saccharomyces cerevisiae IGPD (At_IGPD and Sc_IGPD, respectively). Due to the high flexibility of this region, we performed a total of 3.4 μs of accelerated molecular dynamics simulation to enhance sampling. Inhibitor C348 in At-IGPD exhibited instability in the later stage of simulation, while the characteristic sequence in Sc_IGPD reduced solvent interference and significantly restrained the interaction mode. For the C-Loop-assisted ligand-binding process, we proposed a "Lock-Lid" model. Meanwhile, the dissociated ligand in At_IGPD served as a probe, a metastable pocket was determined at the root of C-Loop, and its rationality was proved by theoretical verification and enzyme mutation experiments. This study complemented the important structural features of C-Loop and provided a basis for the design of selective inhibitors. Considering the absence in mammals, we suggested that IGPD could be a promising germicide target.