| 2014 |
TMEM163 physically interacts with TRPML1 ion channel, and this interaction depends on TMEM163's N-terminus (deletion of part of the N-terminus disrupts it). TRPML1 co-expression reduces plasma membrane levels of TMEM163, while a PM-retained TRPML1 mutant retains TMEM163 at the PM. TMEM163 knockdown or co-knockdown with TRPML1 causes significantly elevated intracellular zinc levels, demonstrating a cooperative role in cellular zinc homeostasis. |
Yeast two-hybrid, co-immunoprecipitation, mass spectrometry, confocal microscopy, N-terminal deletion mutagenesis, fluorescent zinc assays, siRNA knockdown in HEK-293 cells |
Traffic (Copenhagen, Denmark) |
High |
25130899
|
| 2007 |
TMEM163 (SV31) is a synaptic vesicle membrane protein of 31 kDa with six putative transmembrane helices, localized to synaptic vesicles in select brain regions including glutamatergic and GABAergic nerve terminals, as demonstrated by subcellular fractionation, immunocytochemistry of brain sections, and immunoelectron microscopy. |
Subcellular fractionation, heterologous expression, immunocytochemistry of brain sections, immunoelectron microscopy |
Journal of neurochemistry |
High |
17623043
|
| 2011 |
TMEM163 (SV31) binds divalent cations Zn2+ and Ni2+ and to a minor extent Cu2+, but not Fe2+, Co2+, Mn2+, or Ca2+. In PC12 cells, SV31-RFP partially co-fractionates with synaptic-like vesicle markers and early endosome marker Rab5, and localizes near the plasma membrane in a compartment positive for SNAP-25 and syntaxin1A. |
Recombinant protein metal-binding assays, heterologous transfection of PC12 cells, fluorescent zinc indicator (FluoZin-3), sucrose density gradient fractionation, immunocytochemistry |
Journal of neurochemistry |
High |
21668449
|
| 2016 |
TMEM163 (SV31) assembles into dimers when inserted into nanodisc bilayers, with dimerization increasing Zn2+ binding. Site-directed mutagenesis of two conserved aspartate residues markedly reduces Zn2+ binding but does not affect dimerization. Chemical modification of histidine residues also reduces Zn2+ binding. Reconstituted in proteoliposomes, SV31 mediates proton-dependent Zn2+ transport with a Km of 44.3 μM, requiring external Zn2+ and internal acidic pH. |
Cell-free protein synthesis, nanodisc reconstitution, site-directed mutagenesis, native mass spectrometry, proteoliposome transport assay with FluoZin-1 |
Journal of neurochemistry |
High |
27917477
|
| 2019 |
Human TMEM163 functions as a zinc efflux transporter in cells. Cells stably or transiently expressing TMEM163 show significantly reduced intracellular zinc levels as measured by two fluorescent zinc dyes and radionuclide 65Zn. Alanine substitution of two conserved aspartate residues (D124A/D128A) or the E286K variant significantly reduces zinc efflux, establishing these residues as important for transport function. Phylogenetic analysis places TMEM163 within the CDF/SLC30 zinc efflux family. |
Stable and transient expression in human cell lines, fluorescent zinc dyes (two independent), 65Zn radionuclide assay, TPEN chelation control, site-directed mutagenesis, sequence alignment and phylogenetics |
Archives of biochemistry and biophysics |
High |
31697912
|
| 2019 |
Knockdown of endogenous Tmem163 in MIN6 pancreatic beta cells results in increased intracellular zinc, increased total insulin content, but compromised glucose-stimulated insulin secretion and enhanced cellular glucose uptake, placing TMEM163 in the pathway of zinc-regulated insulin storage and secretion. |
siRNA knockdown in MIN6 cells, intracellular zinc measurement, insulin content and secretion assays, glucose uptake assay |
Biochemical and biophysical research communications |
Medium |
31813547
|
| 2020 |
TMEM163 specifically modulates the channel properties and pharmacology of ATP-gated P2X receptors (P2XRs) in vivo. Genome-wide ORF screening identified TMEM163 as a P2XR modulator; it is required for full function of neuronal P2XR and pain-related ATP-evoked behavior. |
Genome-wide ORF functional screen, electrophysiology (implied by channel property assessment), in vivo behavioral assays for pain |
Cell reports |
Medium |
32492420
|
| 2022 |
Heterozygous missense variants in TMEM163 residing in the conserved cytoplasmic domain cause hypomyelinating leukodystrophy. Functional in vitro analysis shows significant impairment of zinc efflux by mutant proteins. Expression of mutant TMEM163 in an oligodendroglial cell line reduces mRNA expression of key myelin genes, decreases branching, and increases cell death, establishing a role for TMEM163-mediated zinc efflux in oligodendrocyte development. |
Whole exome sequencing, in vitro zinc efflux assays with disease variants, oligodendroglial cell line expression, qPCR for myelin genes, cell morphology and viability assays |
Brain : a journal of neurology |
High |
35953447
|
| 2022 |
Two de novo TMEM163 variants (L76R, L76P) cause hypomyelinating leukodystrophy. Functional zinc flux assays show L76R attenuates and L76P enhances zinc efflux. In zebrafish, knockdown of tmem163a/b causes myelin deficit, locomotor disability, and oligodendrocyte/neuron loss; wild-type human TMEM163 rescues the morphant phenotype, while L76R and L76P mutants fail to rescue or worsen it. TMEM163 localizes to the plasma membrane, lysosomes, early endosomes, and other vesicular compartments. |
Functional zinc flux assays (HeLa stable expression), zebrafish morphant model (siRNA knockdown of tmem163a/b), mRNA rescue experiments, locomotor assays, immunostaining for myelin and oligodendrocyte markers |
Cells |
High |
35455965
|
| 2022 |
TMEM163 forms homodimers and heterodimerizes with ZNT1, ZNT2, and ZNT3, but not with ZNT4 (negative control for heterodimerization was not excluded). TMEM163 and ZNT proteins partially co-localize in cells. TMEM163/ZNT heterodimers exhibit similar zinc efflux function as TMEM163 homodimers. Plasma membrane localization of TMEM163 is not markedly altered by ZNT co-expression. |
Co-immunoprecipitation with unique peptide tags, Western blot, confocal microscopy co-localization, cell surface biotinylation, functional zinc flux assays (FluoZin-3, Newport Green) |
Biochemistry and biophysics reports |
Medium |
36204728
|
| 2024 |
TMEM163 is expressed in a subset of dorsal root ganglion neurons enriched with vesicular Zn2+. These neurons form direct synapses with spinal NPY+ inhibitory interneurons and modulate their activity. In aged mice, TMEM163 expression and vesicular Zn2+ concentration are elevated in central terminals of TMEM163+ afferents; excessive vesicular Zn2+ release dampens NPY+ interneuron activity, causing disinhibition of itch circuits and chronic itch. Zinc chelation in the spinal dorsal horn relieves itch in aged mice. |
Immunofluorescence, fluorescent zinc imaging, conditional knockout/neuron-specific analysis, electrophysiology of spinal interneurons, zinc chelation pharmacology in vivo (aged mice) |
PLoS biology |
Medium |
39602426
|
| 2025 |
ZnT3 and TMEM163 physically interact (co-immunoprecipitation confirmed) and cooperatively regulate zinc homeostasis in hippocampal neurons. Under oxygen-glucose deprivation (OGD), both proteins are upregulated and translocate from the cell membrane to the cytoplasm. Overexpression of TMEM163 exacerbates extracellular zinc efflux and neuronal apoptosis under OGD; silencing attenuates zinc overload and neurodegeneration. TMEM163 thus contributes to ischemia-induced neuronal injury via zinc dyshomeostasis. |
siRNA silencing and plasmid overexpression in rat primary hippocampal neurons, OGD model, MTT assay, TUNEL staining, FluoZin-3 zinc measurement, ELISA, co-immunoprecipitation, cell surface biotinylation, RT-qPCR, Western blot |
Frontiers in bioscience (Landmark edition) |
Medium |
41504061
|
| 2026 |
A conserved N-terminal acidic dileucine motif (LEDRGL69L70) in TMEM163 is essential for interaction with BLOC-1 and AP-3 complexes but dispensable for binding AP-1, AP-2, and BLOC-2. Mutation of this motif causes TMEM163 accumulation at the plasma membrane. Loss of BLOC-1 or AP-3 enhances TMEM163 binding to the other complex and results in differential abnormal endo-lysosomal localization, indicating competitive binding and sequential sorting of TMEM163 by AP-3 and BLOC-1 to platelet dense granules. TMEM163 is degraded via the proteasome in cells lacking AP-1, AP-2, AP-3, BLOC-1, or BLOC-2. |
Co-immunoprecipitation, mutagenesis of dileucine motif, confocal microscopy subcellular localization, MEG-01 cell lines deficient in endosomal trafficking complexes, proteasome inhibition assays, cell surface biotinylation |
The Journal of biological chemistry |
High |
41985787
|