A5088758531- Ion channel regulation and function
- Cardiac electrophysiology and arrhythmias
- Plant Stress Responses and Tolerance
- Plant biochemistry and biosynthesis
- Neuroscience and Neuropharmacology Research
- Enzyme Structure and Function
- Glycosylation and Glycoproteins Research
- Biochemical and Molecular Research
- Microbial Natural Products and Biosynthesis
- Ubiquitin and proteasome pathways
- Ion Transport and Channel Regulation
- Neuroscience and Neural Engineering
- Plant nutrient uptake and metabolism
- Mass Spectrometry Techniques and Applications
- Protein Structure and Dynamics
- RNA and protein synthesis mechanisms
- Lipid metabolism and biosynthesis
- Erythrocyte Function and Pathophysiology
- Mitochondrial Function and Pathology
- Skin and Cellular Biology Research
- Genetics and Neurodevelopmental Disorders
- Cell Adhesion Molecules Research
- RNA Research and Splicing
- Molecular Sensors and Ion Detection
- Plant Molecular Biology Research
Tel Aviv University
2016-2025
Tel Aviv Sourasky Medical Center
2016-2024
Israel Defense Forces Medical Corps
2022
Sheba Medical Center
2022
University of Haifa
1985
Epidermolysis bullosa simplex (EBS) refers to a heterogeneous group of inherited skin disorders characterized by blister formation within the basal cell layer. The disease is marked variations in phenotype severity, suggesting co-inheritance genetic modifiers. We identified three deleterious variants HMCN1 that co-segregated with more severe 20 individuals EBS caused mutations KRT14, encoding keratin 14 (K14). codes for hemicentin-1. Protein modeling, molecular dynamics simulations, and...
Significance Na + /Ca 2+ exchangers (NCXs) have a key role in the homeostasis of cellular Ca and consequently are implicated diverse human-health disorders, including neurodegenerative cardiovascular diseases. A detailed understanding molecular mechanisms these membrane proteins is therefore interest from fundamental biomedical standpoints. Here, we establish structural mechanism recognition prokaryotic NCX homolog, using atomistic molecular-dynamics simulations based on recently reported...
The cardiac Na+/Ca2+ exchanger (NCX) regulates cellular [Ca2+]i and plays a central role in health disease, but its molecular regulation is poorly understood. Here we report on how protons affect this electrogenic transporter by modulating two critically important NCX C2 regulatory domains, Ca2+ binding domain-1 (CBD1) CBD2. transport rate intact ventricular myocytes was measured as membrane current, INCX, whereas [H+]i varied using an ammonium chloride "rebound" method at constant...
Mutations in the SCN8A gene, encoding voltage-gated sodium channel Na
In NCX proteins CBD1 and CBD2 domains are connected through a short linker (3 or 4 amino acids) forming regulatory tandem (CBD12). Only three of the six CBD12 Ca(2+)-binding sites contribute to regulation. Two them located on (K(d) = approximately 0.2 microM), one is 5 microM). Here we analyze how intrinsic properties individual affected by linker-dependent interactions in (AD splice variant). The + have comparable K(d) values but differ dramatically their Ca(2+) dissociation kinetics....
Na+/Ca2+ exchanger (NCX) proteins mediate Ca2+-fluxes across the cell membrane to maintain Ca2+ homeostasis in many types. Eukaryotic NCX contains Ca2+-binding regulatory domains, CBD1 and CBD2. binding a primary sensor (Ca3-Ca4 sites) on activates mammalian NCXs, whereas CALX, Drosophila ortholog, displays an inhibitory response Ca2+. To further elucidate underlying mechanisms, we determined 2.7 Å crystal structure of CBD12-E454K, two-domain construct that retains wild-type properties. In...
Abstract In analogy with many other proteins, Na + /Ca 2+ exchangers (NCX) adapt an inverted twofold symmetry of repeated structural elements, while exhibiting a functional asymmetry by stabilizing outward-facing conformation. Here, structure-based mutant analyses the Methanococcus jannaschii exchanger (NCX_Mj) were performed in conjunction HDX-MS (hydrogen/deuterium exchange mass spectrometry) to identify structure-dynamic determinants asymmetry. identified hallmark differences backbone...
Abstract Objective Dravet syndrome (Dravet) is a severe childhood epileptic encephalopathy. The disease begins with febrile stage, characterized by seizures otherwise normal development. Progression to the worsening stage features recurrent intractable and presentation of additional nonepileptic comorbidities, including global developmental delay, hyperactivity, motor deficits. Later in life, at stabilization seizure burden decreases, whereas Dravet‐associated comorbidities persist. To date,...
Abstract The human cis -prenyltransferase (h -PT) is an enzymatic complex essential for protein N-glycosylation. Synthesizing the precursor of glycosyl carrier dolichol-phosphate, mutations in h -PT cause severe diseases. Here, we reveal that exhibits a heterotetrameric assembly solution, consisting two catalytic dehydrodolichyl diphosphate synthase (DHDDS) and inactive Nogo-B receptor (NgBR) heterodimers. Importantly, 2.3 Å crystal structure reveals tetramer assembles via DHDDS C-termini as...
Abstract Chloride Intracellular Channel (CLIC) family members uniquely transition between soluble and membrane-associated conformations. Despite decades of extensive functional structural studies, CLICs’ function as ion channels remains debated, rendering our understanding their physiological role incomplete. Here, we expose the CLIC5 a fusogen. We demonstrate that purified directly interacts with membrane induces fusion, reflected by increased liposomal diameter lipid content mixing...
Summary Whereas tetrahydrofolate is an essential cofactor in all bacteria, the gene that encodes enzyme dihydrofolate reductase (DHFR) could not be identified many of bacteria whose genomes have been entirely sequenced. In this communication we show halophilic archaea Halobacterium salinarum and Haloarcula marismortui contain genes coding for proteins with N‐terminal domain homologous to synthase (FolC) a C‐terminal dihydropteroate (FolP). These are able complement Haloferax volcanii mutant...
In eukaryotic Na(+)/Ca(2+) exchangers (NCX) the Ca(2+) binding CBD1 and CBD2 domains form a two-domain regulatory tandem (CBD12). An allosteric sensor (Ca3-Ca4 sites) is located on CBD1, whereas contains splice-variant segment. Recently, Ca(2+)-driven interdomain switch has been described, albeit how it couples with signal propagation remains unclear. To resolve dynamic features of Ca(2+)-induced conformational transitions we analyze here distinct splice variants mutants isolated CBD12 at...
Members of the Ca(2+)/cation exchanger superfamily (Ca(2+)/CA) share structural similarities (including highly conserved ion-coordinating residues) while exhibiting differential selectivity for Ca(2+), Na(+), H(+), K(+), and Li(+). The archaeal Na(+)/Ca(2+) (NCX_Mj) its mammalian orthologs are selective whereas mitochondrial ortholog (NCLX) can transport either Li(+) or Na(+) in exchange with Ca(2+). Here, structure-based replacement residues NCX_Mj resulted a capacity transporting Li(+),...
The Ca2+-dependent allosteric regulation of Na+/Ca2+ exchanger (NCX) proteins represents Ca2+ interaction with the cytosolic domains, CBD1 (calcium-binding domain 1) and CBD2, which is associated either activation, inhibition or no response to regulatory in a given splice variant. contains high affinity Ca2+-sensor (which highly conserved among variants), whereas primary information upon binding modified by alternative splicing yielding diverse responses Ca2+. To resolve structure-dynamic...
Mammalian Na+/Ca2+ exchangers, NCX1 and NCX3, generate splice variants, whereas NCX2 does not. The CBD1 CBD2 domains form a regulatory tandem (CBD12), where Ca2+ binding to activates (bearing the splicing segment) alleviates Na+-induced inactivation. Here, NCX2-CBD12, NCX3-CBD12-B, NCX3-CBD12-AC proteins were analyzed by small-angle X-ray scattering (SAXS) hydrogen-deuterium exchange mass-spectrometry (HDX-MS) resolve variances in NCX3 variants. SAXS revealed unified model, according which...