A Preliminary Analysis of the Effects of NF058 on Activities of Neuronal and Inducible Isoforms of Nitric Oxide Synthase (NOS)
Diffenderfer, Katherine M.
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Nitric oxide synthase has become the target of much research due to the apparent link of its product nitric oxide (•NO) in neurodegenerative diseases such as Parkinson's and Alzheimer's disease. The pathology of •NO production is believed to be the result of activity of either nNOS or iNOS in the brain. Thus, ways to selectively inhibit the isoforms could prove useful for therapy. The calmodulin binding sequence within the various NOS isoforms offers an ideal drug design target site, as these sites are varied. Suramin and one of its analogs (NF307) has been demonstrated to distinguish among calmodulin binding sequences. Given the differences in the CaM-binding sequences of NOSs, the ability of a suramin analog, NF058, to select between two NOS isoforms has been studied. Following E. coli expression, column purification, and activity analyses (hemoglobin and cytochrome c assays), it was demonstrated that NF058 selectively inhibits the neuronal NOS isoform, with a K| of 4.27 u.M. The K| for inducible NOS need concentrations greater than 94 |iM to be determined. In addition, NF058 was demonstrated to be a mixed inhibitor of neuronal NOS, though its mechanism of inhibition still needs to be determined. Thus, with further research on the mechanism of inhibition by NF058, possible drug compounds could be synthesized, using NF058 as the lead compound in neurodegenerative drug-therapy treatments.