Identification of small molecule compounds with higher binding affinity to guanine deaminase (cypin) than guanine.
Guanine deaminase (GDA; cypin) is an important metalloenzyme that processes the first step in purine catabolism, converting guanine to xanthine by hydrolytic deamination. In higher eukaryotes, GDA also plays an important role in the development of neuronal morphology by regulating dendritic arborization. In addition to its role in the maturing brain, GDA is thought to be involved in proper liver function since increased levels of GDA activity have been correlated with liver disease and transplant rejection. Although mammalian GDA is an attractive and potential drug target for treatment of both liver diseases and cognitive disorders, prospective novel inhibitors and/or activators of this enzyme have not been actively pursued. In this study, we employed the combination of protein structure analysis and experimental kinetic studies to seek novel potential ligands for human guanine deaminase. Using virtual screening and biochemical analysis, we identified common small molecule compounds that demonstrate a higher binding affinity to GDA than does guanine. In vitro analysis demonstrates that these compounds inhibit guanine deamination, and more surprisingly, affect GDA (cypin)-mediated microtubule assembly. The results in this study provide evidence that an in silico drug discovery strategy coupled with in vitro validation assays can be successfully implemented to discover compounds that may possess therapeutic value for the treatment of diseases and disorders where GDA activity is abnormal.
Fernández
José R
JR
Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, 604 Allison Road, Piscataway, NJ 08854-8082, USA.
Sweet
Eric S
ES
Welsh
William J
WJ
Firestein
Bonnie L
BL
D001665:Binding Sites; D003072:Cognition Disorders / Q000188:drug therapy; D004734:Energy Metabolism; D006147:Guanine / Q000378:metabolism*; D006148:Guanine Deaminase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008107:Liver Diseases / Q000188:drug therapy; D008870:Microtubules / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000627:therapeutic use; D013329:Structure-Activity Relationship
D008024:Ligands; D054852:Small Molecule Libraries; D006147:Guanine; D006148:Guanine Deaminase