EXPLORING THE DIVERSE APPLICATIONS OF X-RAY CRYSTALLOGRAPHY: ITS SIGNIFICANCE IN STRUCTURAL BIOLOGY, BIOCHEMISTRY, AND MEDICINAL CHEMISTRY

X-ray crystallography has brought about a revolution in the fields of structural biology, biochemistry, and medicinal chemistry. By providing atomic-level details, this method has significantly advanced our comprehension of biological processes and facilitated the development of effective therapeutics. At the Center for the Research and Advancement of Fragments and Molecular Targets (CRAFT), we extensively employ X-ray crystallography in conjunction with other biophysical techniques to characterize macromolecular targets and propel drug discovery endeavors. One of our key applications involves the utilization of X-ray crystallography fragment screening (XCFS) to identify binding sites in drug targets. Notably, our work on human dihydroorotate dehydrogenase, a target for broad-spectrum antiviral drugs, has resulted in the identification of critical hotspots and the development of active hit compounds against SARS-CoV-2. Similarly, in our pursuit of antileishmaniasis drugs within medicinal chemistry, crystal structures of Leishmania braziliensis dihydroorotate dehydrogenase complexed with barbituric acid-based inhibitors have offered a molecular foundation for designing a new generation of compounds with efficacy against leishmania. Moreover, our research endeavors have validated the efficacy of a novel technique called MIDAS, which relies on X-ray crystallography. MIDAS enables the sensitive identification of protein-metabolite interactions. Through the analysis of enzymes from central carbon metabolism, we have successfully identified 830 protein-metabolite interactions, including previously undiscovered and unexpected interactions such as the binding of human fumarase with 2-amino-3-phosphonopropionic acid (AP-3), a component of phosphonate metabolism.
With continuous advancements, X-ray crystallography holds tremendous promise for further breakthroughs in these disciplines. At CRAFT, we remain dedicated to contributing to the progress of X-ray crystallography and its wide-ranging applications, with the ultimate goal of driving innovation and augmenting our comprehension of intricate biological systems.

This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP # 2020/06190-0, # 2008/08262-6), by CNPq (BRICS STI COVID-19 #441038/2020-4) and by National Institutes of Health (NIH #R01 R01 AI160379).