Interaction of dynamin I with NAP-22, a neuronal protein enriched in the presynaptic region. (14th May 2018)

• Record Type: Journal Article
• Authors: Ueno, Satoko; Miyoshi, Hiroshi; Maruyama, Yoko; Morita, Mitsuhiro; Maekawa, Shohei
• Is Part Of: Neuroscience letters. Volume 675(2018)
• Page Start: 59
• Page End: 63
• Publication Date: 2018-05-14
• Subjects: DRM detergent-resistant low-density membrane microdomain fraction -- CaM calmodulin -- TME solution 10 mM Tris-HCl, 1 mM MgCl2, 1 mM EGTA, pH 7.4 -- PMSF phenylmethylsulfonyl fluoride -- AS affinity screening -- AS buffer 10 mM Tris-HCl, 1 mM MgCl2, 0.2 mM EGTA, 140 mM NaCl, pH 7.4 -- SAS saturated ammonium sulfate -- PS phosphatidylserine -- PIP2 phosphatidylinositol 4, 5-bisphosphate; Dynamin -- NAP-22 -- Calmodulin -- Phosphatidylserine -- Endocytosis; Neurology -- Periodicals; Neurology -- Periodicals; Research -- Periodicals; Neurologie -- Périodiques; Neuroanatomie -- Périodiques; Neuropharmacologie -- Périodiques; Neurophysiologie -- Périodiques; Neurology; Periodicals; Electronic journals; 617.48
• DOI: 10.1016/j.neulet.2018.03.061
• Languages: English
• ISSNs: 0304-3940

Abstract

Abstract: Neurons have well-developed membrane microdomains called "rafts" that are recovered as a detergent-resistant low-density membrane microdomain fraction (DRM). NAP-22 is one of the major protein components of neuronal DRM and localizes in the presynaptic region. In order to know the role of NAP-22 in the synaptic transmission, NAP-22 binding proteins in the cytosol were searched with an affinity screening with NAP-22 as a bait and several protein bands were detected. Using mass-analysis and western blotting, one of the main band of ∼90 kDa was identified as dynamin I. The GTPase activity of dynamin I was partly inhibited by NAP-22 expressed in bacteria and this inhibition was recovered by the addition of calmodulin, a NAP-22 binding protein. The GTPase activity of dynamin was known to be activated with acidic membrane lipids such as phosphatidylserine and the addition of NAP-22, a phosphatidylserine binding protein, inhibited the activation of the GTPase by this lipid. Since NAP-22 localizes on the presynaptic plasma membrane and on synaptic vesicles, these results suggest the participation of NAP-22 in the membrane cycling through binding to dynamin and acidic membrane lipids at the presynaptic region.