S-Nitrosylation of Divalent Metal Transporter 1 Enhances Iron Uptake to Mediate Loss of Dopaminergic Neurons and Motoric Deficit.

Autor: Liu C; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Zhang CW; Key Laboratory of Flexible Electronics (KLOFE) and Institute of Advanced Materials (IAM), Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing Tech University, Nanjing 211816, People's Republic of China.; Department of Research, National Neuroscience Institute, Singapore 308433., Lo SQ; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Ang ST; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Chew KCM; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.; Department of Research, National Neuroscience Institute, Singapore 308433., Yu D; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Chai BH; Department of Research, National Neuroscience Institute, Singapore 308433., Tan B; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Tsang F; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.; Department of Research, National Neuroscience Institute, Singapore 308433., Tai YK; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.; Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119228., Tan BWQ; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Liang MC; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Tan HT; Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117596., Tang JY; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597., Lai MKP; Department of Pharmacology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599., Chua JJE; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.; Neurobiology and Ageing Programme, Life Sciences Institute, Centre for Life Sciences, National University of Singapore, Singapore 117456.; Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore 138673., Chung MCM; Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117596., Khanna S; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.; Neurobiology and Ageing Programme, Life Sciences Institute, Centre for Life Sciences, National University of Singapore, Singapore 117456., Lim KL; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.; Department of Research, National Neuroscience Institute, Singapore 308433.; Neurobiology and Ageing Programme, Life Sciences Institute, Centre for Life Sciences, National University of Singapore, Singapore 117456.; Neuroscience and Behavioral Disorders Program, Duke-NUS Medical School, Singapore 169857, and., Soong TW; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, phsstw@nus.edu.sg.; Department of Research, National Neuroscience Institute, Singapore 308433.; Neurobiology and Ageing Programme, Life Sciences Institute, Centre for Life Sciences, National University of Singapore, Singapore 117456.; NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore 117456.
Jazyk: angličtina
Zdroj: The Journal of neuroscience : the official journal of the Society for Neuroscience [J Neurosci] 2018 Sep 26; Vol. 38 (39), pp. 8364-8377. Date of Electronic Publication: 2018 Aug 13.
DOI: 10.1523/JNEUROSCI.3262-17.2018
Abstrakt: 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 Fe 2+ 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 Mn 2+ or Fe 2+ 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 evaluate in vivo significance, 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 Fe 2+ deposition, JNK activation, dopaminergic neuronal loss and deficit in motoric activity, and these were rescued by the NO synthase inhibitor l-NAME or by the DMT1-selective blocker ebselen. Lentiviral knockdown of DMT1 abolished LPS-induced dopaminergic neuron loss. SIGNIFICANCE STATEMENT Neuroinflammation and high cytoplasmic Fe 2+ 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 Fe 2+ 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 using l-NAME, a NO synthase inhibitor, or by ebselen, a DMT1-selective blocker.
(Copyright © 2018 the authors 0270-6474/18/388365-14$15.00/0.)
Databáze: MEDLINE
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