S-Nitrosylation of Divalent Metal Transporter 1 Enhances Iron Uptake to Mediate Loss of Dopaminergic Neurons and Motoric Deficit
| Autor: | Katherine C. M. Chew, Seok Ting Ang, Sanjay Khanna, Bryce W.Q. Tan, Fai Tsang, Chao Liu, Chengwu Zhang, Bobby Tan, Mui Cheng Liang, Dejie Yu, Hwee Tong Tan, Tuck Wah Soong, John Jia En Chua, Yee Kit Tai, Bing Han Chai, Shun Qiang Lo, Jia Ying Tang, Kah-Leong Lim, Mitchell K.P. Lai, Maxey C. M. Chung |
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| Rok vydání: | 2018 |
| Předmět: |
Lipopolysaccharides
Male 0301 basic medicine Iron Mice Transgenic Substantia nigra Pharmacology Nitric Oxide Nitric oxide 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Animals Humans Cation Transport Proteins Research Articles Neuroinflammation Inflammation biology Ebselen Dopaminergic Neurons General Neuroscience digestive oral and skin physiology Dopaminergic Parkinson Disease Transporter DMT1 S-Nitrosylation Substantia Nigra 030104 developmental biology chemistry biology.protein Female Erratum Locomotion 030217 neurology & neurosurgery |
| Zdroj: | J Neurosci |
| ISSN: | 1529-2401 0270-6474 |
| Popis: | Elevated iron deposition has been reported in Parkinson's disease (PD). However, the route of iron uptake leading to high deposition in the substantia nigra is unresolved. Here, we show a mechanism in enhanced Fe2+uptake via S-nitrosylation of divalent metal transporter 1 (DMT1). While DMT1 could be S-nitrosylated by exogenous nitric oxide donors, in human PD brains, endogenously S-nitrosylated DMT1 was detected in postmortem substantia nigra. Patch-clamp electrophysiological recordings and iron uptake assays confirmed increased Mn2+or Fe2+uptake through S-nitrosylated DMT1. We identified two major S-nitrosylation sites, C23 and C540, by mass spectrometry, and DMT1 C23A or C540A substitutions abolished nitric oxide (NO)-mediated DMT1 current increase. To evaluatein vivosignificance, lipopolysaccharide (LPS) was stereotaxically injected into the substantia nigra of female and male mice to induce inflammation and production of NO. The intranigral LPS injection resulted in corresponding increase in Fe2+deposition, JNK activation, dopaminergic neuronal loss and deficit in motoric activity, and these were rescued by the NO synthase inhibitorl-NAME or by the DMT1-selective blocker ebselen. Lentiviral knockdown of DMT1 abolished LPS-induced dopaminergic neuron loss.SIGNIFICANCE STATEMENTNeuroinflammation and high cytoplasmic Fe2+levels have been implicated in the initiation and progression of neurodegenerative diseases. Here, we report the unexpected enhancement of the functional activity of transmembrane divalent metal transporter 1 (DMT1) by S-nitrosylation. We demonstrated that S-nitrosylation increased DMT1-mediated Fe2+uptake, and two cysteines were identified by mass spectrometry to be the sites for S-nitrosylation and for enhanced iron uptake. One conceptual advance is that while DMT1 activity could be increased by external acidification because the gating of the DMT1 transporter is proton motive, we discovered that DMT1 activity could also be enhanced by S-nitrosylation. Significantly, lipopolysaccharide-induced nitric oxide (NO)-mediated neuronal death in the substantia nigra could be ameliorated by usingl-NAME, a NO synthase inhibitor, or by ebselen, a DMT1-selective blocker. |
| Databáze: | OpenAIRE |
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