A5021319752- Botulinum Toxin and Related Neurological Disorders
- Neurological disorders and treatments
- Neuroscience and Neuropharmacology Research
- Cellular transport and secretion
- Hereditary Neurological Disorders
- Biochemical and Structural Characterization
- Nicotinic Acetylcholine Receptors Study
- Ion channel regulation and function
- Marine Toxins and Detection Methods
- Monoclonal and Polyclonal Antibodies Research
- Toxin Mechanisms and Immunotoxins
- Transgenic Plants and Applications
- Cholinesterase and Neurodegenerative Diseases
- Neurotransmitter Receptor Influence on Behavior
- Atherosclerosis and Cardiovascular Diseases
- Neurobiology and Insect Physiology Research
- Receptor Mechanisms and Signaling
- Glycosylation and Glycoproteins Research
- Protease and Inhibitor Mechanisms
- RNA Interference and Gene Delivery
- Venomous Animal Envenomation and Studies
- Lipoproteins and Cardiovascular Health
- Cell Adhesion Molecules Research
- Biotin and Related Studies
- Ubiquitin and proteasome pathways
New York University
2013-2024
CytoDel (United States)
2014
The University of Texas Southwestern Medical Center
1995-2002
Howard Hughes Medical Institute
1994-1999
Max Planck Institute of Experimental Medicine
1999
Columbia University Irving Medical Center
1998
Osaka University
1997
Science and Technology Corporation (United States)
1997
Laboratory of Molecular Genetics
1997
PSD-95 is a component of postsynaptic densities in central synapses. It contains three PDZ domains that localize N -methyl- d -aspartate receptor subunit 2 (NMDA2 receptor) and K + channels to In mouse forebrain, bound the cytoplasmic COOH-termini neuroligins, which are neuronal cell adhesion molecules interact with β-neurexins form intercellular junctions. Neuroligins bind third domain PSD-95, whereas NMDA2 receptors first second domains. Thus different specialized for distinct functions....
At the synapse, presynaptic membranes specialized for vesicular traffic are linked to postsynaptic signal transduction. The mechanisms that connect pre- and into synaptic junctions unknown. Neuroligins beta-neurexins neuronal cell-surface proteins bind each other form asymmetric intercellular junctions. To test whether neuroligin/beta-neurexin junction is related synapses, we generated characterized monoclonal antibodies neuroligin 1. With these antibodies, show 1 synaptic. localization,...
The low-density lipoprotein receptor mediates cholesterol homeostasis through endocytosis of lipoproteins. It discharges its ligand in the endosome at pH < 6. In crystal structure = 5.3, ligand-binding domain (modules R2 to R7) folds back as an arc over epidermal growth factor precursor homology (the modules A, B, β propeller, and C). R4 R5, which are critical for binding, associate with propeller via their calcium-binding loop. We propose a mechanism release whereby functions alternate...
Neuroligin 1 is a neuronal cell surface protein that binds to subset of neurexins, polymorphic proteins are also localized on neurons (Ichtchenko, K., Hata, Y., Nguyen, T., Ullrich, B., Missler, M., Moomaw, C., and Südhof, T. C. (1995) Cell 81, 435-443). We now describe two novel neuroligins called 2 3 similar in structure sequence neuroligin 1. All contain an N-terminal hydrophobic with the characteristics cleaved signal peptide followed by large esterase homology domain, highly conserved...
α-Latrotoxin, a potent excitatory neurotoxin, binds to two receptors: G-protein-coupled receptor calledCIRL/latrophilin 1 (CL1) and cell-surface protein called neurexin Iα. We now show that CL1 belongs family of closely related receptors CL1, CL2, CL3. CLs exhibit an unusual multidomain structure with similar alternative splicing large extra- intracellular sequences. share domains other receptors, lectins, olfactomedins/myocilin. In addition, contain novel, widespread cysteine-rich domain...
Synaptotagmin I is a Ca2+-binding protein of synaptic vesicles that serves as Ca2+ sensor for neurotransmitter release and was the first member found large family trafficking proteins. We have now identified novel synaptotagmin, synaptotagmin XI, highly expressed in brain at lower levels other tissues. Like synaptotagmins, XI has single transmembrane region two cytoplasmic C2-domains but most closely related to IV with which it forms new subclass synaptotagmins. The C2-domain (the...
Neurexins, a family of neuronal cell-surface proteins, consist the longer a-neurexins (Ia, IIa, and IIIa) shorter p-neurexins (Ip IIp) with identical C termini but distinct N termini.a-Neurexins have structure cell surface receptors, membrane topology conservation is unknown.W e now characterized cDNA clones encoding bovine neurexins IS IIIp, thereby demonstrating presence p-form for neurexin 111 evolutionary in mammals.Similar to a-neurexins, pneurexins were found be highly O-glycosylated...
The calcium-independent receptor of α-latrotoxin (CIRL), a neuronal cell surface implicated in the regulation exocytosis, is natural chimera adhesion protein and G protein-coupled (GPCR). In contrast with canonic GPCRs, CIRL consists two heterologous non-covalently bound subunits, p120 p85, due to endogenous proteolytic processing precursor endoplasmic reticulum. Extracellularly oriented contains hydrophilic domains, whereas p85 resembles generic GPCR. We determined that site cleavage...
Poisoning with alpha-latrotoxin, a neurotoxic protein from black widow spider venom, results in robust increase of spontaneous synaptic transmission and subsequent degeneration affected nerve terminals. The action alpha-latrotoxin involves extracellular binding to its high affinity receptors as first step. One these proteins, CIRL, is neuronal G-protein-coupled receptor implicated the regulation secretion. We now demonstrate that CIRL has two close homologs similar domain structure degree...
Somatodendritic dopamine (DA) release from midbrain DA neurons activates D2 autoreceptors on these cells to regulate their activity. However, the source of autoregulatory remains controversial. Here, we test hypothesis that a given neuron in substantia nigra pars compacta (SNc) are activated primarily by released same cell, rather than its neighbors. Voltage-clamp recording allows monitoring evoked D2-receptor-mediated inhibitory currents (D2ICs) SNc as an index release. Single-cell...
Intraneuronal delivery of a therapeutic, single-domain antibody reverses botulism symptoms and increases survival mice, guinea pigs, monkeys.
Botulinum neurotoxins (BoNTs) are among the deadliest of bacterial toxins. BoNT serotype A and B in particular pose most serious threat to humans because their high potency persistence. To date, there is no effective treatment for late post-exposure therapy botulism patients. Here, we aim develop single-domain variable heavy-chain (VHH) antibodies targeting protease domains (also known as light chain, LC) BoNT/A BoNT/B antidotes post-intoxication treatments. Using a combination X-ray...
Receptor-like protein-tyrosine phosphatase sigma (PTPς) is essential for neuronal development and function. Here we report that PTPς a target of α-latrotoxin, strong stimulator exocytosis. α-Latrotoxin binds to the cell adhesion-like extracellular region PTPς. This binding results in stimulation The toxin-binding site located C-terminal part ectodomain includes two fibronectin type III repeats. intracellular catalytic domains are not required α-latrotoxin secretory response triggered by...
Abstract Botulinum neurotoxin (BoNT) binds to and internalizes its light chain into presynaptic compartments with exquisite specificity. While the native toxin is extremely lethal, bioengineering of BoNT has potential eliminate toxicity without disrupting neuron-specific targeting, thereby creating a molecular vehicle capable delivering therapeutic cargo neuronal cytosol. Building upon previous work, we have developed an toxic d erivative ( ad ) BoNT/C1 through rationally designed amino acid...
Botulinum neurotoxin (BoNT) serotype E is one of three serotypes that cause the preponderance human botulism cases and a Tier 1 Select Agent. BoNT/E unusual among BoNT for its rapid onset short duration intoxication. Here we report two large panels unique, unrelated camelid single-domain antibodies (VHHs) were selected their ability to bind holotoxin and/or light chain protease domain (LC/E). The 19 VHHs which characterized subunit specificity 8 displayed neutralize intoxication neurons....
While covalent drug discovery is reemerging as an important route to small-molecule therapeutic leads, strategies for the and engineering of protein-based irreversible binding agents remain limited. Here, we describe use yeast display in combination with noncanonical amino acids (ncAAs) identify variants single-domain antibodies (sdAbs), also called VHHs nanobodies, targeting botulinum neurotoxin light chain A (LC/A). Starting from a series previously described, structurally characterized...
We have previously described genetic constructs and expression systems that enable facile production of recombinant derivatives botulinum neurotoxins (BoNTs) retain the structural trafficking properties wt BoNTs. In this report we describe one such derivative, BoNT/A ad, which was rendered atoxic by introducing two amino acid mutations to light chain (LC) BoNT/A, is being developed as a molecular vehicle for delivering drugs neuronal cytoplasm. The binding, internalization, intracellular ad...
The incidence of Clostridioides difficile infection (CDI) and associated mortality have increased rapidly worldwide in recent years. Therefore, it is critical to develop new therapies for CDI. Here we report on the development mRNA-LNPs encoding camelid-derived VHH-based neutralizing agents (VNAs) targeting toxins A and/or B C. difficile. In preclinical models, intravenous administration provided serum VNA levels sufficient confer protection mice against severe disease progression following...
Abstract Cyto-012 is a recombinant derivative of Botulinum neurotoxin Type A (BoNT/A). It primarily differs from wild type ( wt ) BoNT/A1 in that it incorporates two amino acid substitutions the catalytic domain light chain (LC) metalloprotease (E 224 > and Y 366 A), designed to provide safer clinical profile. specifically internalized into rat cortical hippocampal neurons, cleaves Synaptosomal-Associated Protein 25 (SNAP-25), substrate BoNT/A, but exhibits slower cleavage kinetics...
Botulinum neurotoxin type A1 (BoNT/A1) is a potent protein toxin responsible for the potentially fatal human illness botulism. Notwithstanding, long-lasting flaccid muscle paralysis caused by BoNT/A has led to its utility as powerful and versatile bio-pharmaceutical. The due specific cleavage of neuronal SNAREs BoNTs. However, actions BoNTs on intoxicated neurons besides have not been studied in detail. In this study we investigated microarray analysis effects catalytically inactive...