The linoleic acid derivative 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoicacid (DCP-LA) has been developed as a promising and novel anti-dementia drug. The present study probed the binding partners of DCP-LA. In the DCP-LA binding assay using rat hippocampal lysates, fluorescein-conjugated DCP-LA produced a fluorescent signal band at 75 kDa. In the matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOFMS) analysis, the signal band corresponded to N-ethylmaleimide-sensitive factor (NSF). In the immunofluorescent cytochemistry using acutely dissociated rat hippocampal neurons, intracellular localization of NSF was well consistent with DCP-LA distribution. The Biacore assay equipped with a sensor chip immobilizing DCP-LA was carried out and the equilibrium dissociation constant (Kd) value between DCP-LA and NSF was measured by applying a variety of concentrations of recombinant NSF. The results of the present study show that DCP-LA binds to NSF specifically, with the Kd value of 3.3x10-7 M.
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