The Benefits of Botulinum Neurotoxin Treatment in a Multitude of Medical Conditions

2018 ◽  
Vol 68 (12) ◽  
pp. 2978-2983
Author(s):  
Tiberiu Paul Neagu ◽  
Ion Cocolos ◽  
Cristian Cobilinschi ◽  
Mirela Tiglis ◽  
Ioan Petre Florescu ◽  
...  
Keyword(s):  
Pain Management ◽  
Membrane Protein ◽  
Botulinum Neurotoxin ◽  
Clinical Applications ◽  
Therapeutic Action ◽  
Presynaptic Membrane ◽  
Attachment Protein ◽  
Protein Receptors ◽  
Botulinum Neurotoxins

Botulinum neurotoxins (BoNTs) are responsible for botulism in humans and vertebrates, being one of the six most catastrophic potential bioterrorism agents. This are ~150 kDa proteins, assembled as a ~50 kDa light chain (LC) and a ~100 kDa heavy chain (HC). The LC acts like a zinc metalloproteinase that cleaves three proteins in neurons, members of the SNARE (Soluble N-ethylmaleimide sensitive fusion attachment protein receptors) family: VAMP (vesicle-associated membrane protein) / synaptobrevin, SNAP-25 (synaptosomal-associated protein 25) and syntaxin. After cleavage of any of this proteins, neurotransmission is blocked and flaccid paralysis of the muscle is installed. This extraordinary restricted tropism for the cholinergic presynaptic membrane makes this drug unique regarding its toxicity, pharmacological and therapeutic use. Taking into consideration the potential of this substance, this paper aims to summarize the most relevant data regarding the mechanism of actions and its main clinical applications, in order to improve medical practice. Therefore, we presented the mechanism of action in order to understand its usage in different pathologies, such as dystonias, spasticity, nephrologic and urologic conditions, cosmetic use, depression, gastroenterologic and proctologic diseases, dermatologic conditions, pathologies specific to plastic surgery and also the role of BoNT therapy in pain management. It is well documented in the literature that important discoveries have been made through recent experimental and clinical studies. Even so, there is still much to learn about the therapeutic action of this drug in terms of molecular and pathophysiological mechanisms, in order to benefit of the whole healing potential of this amazing toxin.

scholarly journals Vesicle-associated membrane protein-2 (synaptobrevin-2) forms a complex with synaptophysin

1995 ◽  
Vol 305 (3) ◽  
pp. 721-724 ◽  
Author(s):  
P Washbourne ◽  
G Schiavo ◽  
C Montecucco
Keyword(s):  
Membrane Protein ◽  
Neurotransmitter Release ◽  
Synaptic Vesicles ◽  
Cross Linking ◽  
Type Ii ◽  
Presynaptic Membrane ◽  
Terminal Part ◽  
Vesicle Docking ◽  
Botulinum Neurotoxins ◽  
Type Ii Membrane Protein

Vesicle-associated membrane protein (VAMP) (or synaptobrevin), a type II membrane protein of small synaptic vesicles, is essential for neuroexocytosis because its proteolysis by tetanus and botulinum neurotoxins types B, D, F and G blocks neurotransmitter release. The addition of cross-linking reagents to isolated small synaptic vesicles induces the formation of 30 and 50 kDa complexes containing the isoform 2 of VAMP (VAMP-2). Whereas the 30 kDa band is a VAMP-2 homodimer, the 50 kDa species results from the cross-linking of VAMP-2 with synaptophysin. This heterodimer also forms in detergent-solubilized vesicles and involves the N-terminal part of VAMP-2. The implications of the existence of a synaptophysin-VAMP-2 complex in the processes of vesicle docking and fusion with the presynaptic membrane are discussed.

scholarly journals Comparative study of commercially available and homemade anti-VAMP7 antibodies using CRISPR/Cas9-depleted HeLa cells and VAMP7 knockout mice

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 1649
Author(s):  
Agathe Verraes ◽  
Beatrice Cholley ◽  
Thierry Galli ◽  
Sebastien Nola
Keyword(s):  
Membrane Protein ◽  
Membrane Fusion ◽  
Hela Cells ◽  
Intracellular Membrane ◽  
Wild Type ◽  
Attachment Protein ◽  
Sensitive Factor ◽  
Protein Receptors ◽  
Brain Extracts ◽  
Intracellular Membrane Fusion

VAMP7 (vesicle-associated membrane protein) belongs to the intracellular membrane fusion SNARE (Soluble N-ethylmaleimide-sensitive factor attachment protein receptors) protein family. In this study, we used CRISPR/Cas9 genome editing technology to generate VAMP7 knockout (KO) human HeLa cells and mouse KO brain extracts in order to test the specificity and the background of a set of commercially available and homemade anti-VAMP7 antibodies. We propose a simple profiling method to analyze western blotting and immunocytochemistry staining profiles and determine the extent of the antibodies’ specificity. Using this method, we were able to rank the performance of a set of available antibodies and further showed an optimized procedure for VAMP7 immunoprecipitation, which we validated using wild-type and KO mouse brain extracts.

scholarly journals Comparative study of commercially available and homemade anti-VAMP7 antibodies using CRISPR/Cas9-depleted HeLa cells and VAMP7 knockout mice

F1000Research ◽  
2019 ◽  
Vol 7 ◽  
pp. 1649 ◽  
Author(s):  
Agathe Verraes ◽  
Beatrice Cholley ◽  
Thierry Galli ◽  
Sebastien Nola
Keyword(s):  
Membrane Protein ◽  
Hela Cells ◽  
Intracellular Membrane ◽  
Wild Type ◽  
Attachment Protein ◽  
Visual Scoring ◽  
Sensitive Factor ◽  
Protein Receptors ◽  
Brain Extracts ◽  
Intracellular Membrane Fusion

VAMP7 (vesicle-associated membrane protein) belongs to the intracellular membrane fusion SNARE (Soluble N-ethylmaleimide-sensitive factor attachment protein receptors) protein family. In this study, we used CRISPR/Cas9 genome editing technology to generate VAMP7 knockout (KO) human HeLa cells and mouse KO brain extracts in order to test the specificity and the background of a set of commercially available and homemade anti-VAMP7 antibodies. We propose a simple profiling method to analyze western blotting and use visual scoring for immunocytochemistry staining to determine the extent of the antibodies’ specificity. Thus, we were able to rank the performance of a set of available antibodies and further showed an optimized procedure for VAMP7 immunoprecipitation, which we validated using wild-type and KO mouse brain extracts.

scholarly journals Phosphorylation of SNAP-23 at Ser95 causes a structural alteration and negatively regulates Fc receptor–mediated phagosome formation and maturation in macrophages

2018 ◽  
Vol 29 (14) ◽  
pp. 1753-1762 ◽  
Author(s):  
Chiye Sakurai ◽  
Makoto Itakura ◽  
Daiki Kinoshita ◽  
Seisuke Arai ◽  
Hitoshi Hashimoto ◽  
...  
Keyword(s):  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Interferon Γ ◽  
Fc Receptor ◽  
Attachment Protein ◽  
Iκb Kinase ◽  
Sensitive Factor ◽  
Protein Receptors ◽  
Fret Efficiency

SNAP-23 is a plasma membrane-localized soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNARE) involved in Fc receptor (FcR)-mediated phagocytosis. However, the regulatory mechanism underlying its function remains elusive. Using phosphorylation-specific antibodies, SNAP-23 was found to be phosphorylated at Ser95 in macrophages. To understand the role of this phosphorylation, we established macrophage lines overexpressing the nonphosphorylatable S95A or the phosphomimicking S95D mutation. The efficiency of phagosome formation and maturation was severely reduced in SNAP-23-S95D–overexpressing cells. To examine whether phosphorylation at Ser95 affected SNAP-23 structure, we constructed intramolecular Förster resonance energy transfer (FRET) probes of SNAP-23 designed to evaluate the approximation of the N termini of the two SNARE motifs. Interestingly, a high FRET efficiency was detected on the membrane when the S95D probe was used, indicating that phosphorylation at Ser95 caused a dynamic structural shift to the closed form. Coexpression of IκB kinase (IKK) 2 enhanced the FRET efficiency of the wild-type probe on the phagosome membrane. Furthermore, the enhanced phagosomal FRET signal in interferon-γ–activated macrophages was largely dependent on IKK2, and this kinase mediated a delay in phagosome-lysosome fusion. These results suggested that SNAP-23 phosphorylation at Ser95 played an important role in the regulation of SNARE-dependent membrane fusion during FcR-mediated phagocytosis.

Role of tethering factors in secretory membrane traffic

2006 ◽  
Vol 290 (1) ◽  
pp. C11-C26 ◽  
Author(s):  
Elizabeth Sztul ◽  
Vladimir Lupashin
Keyword(s):  
Coiled Coil ◽  
Membrane Traffic ◽  
Cellular Responses ◽  
Regulatory Proteins ◽  
Attachment Protein ◽  
Experimental Systems ◽  
Sensitive Factor ◽  
Protein Receptors ◽  
Protein Components

Coiled-coil and multisubunit tethers have emerged as key regulators of membrane traffic and organellar architecture. The restricted subcellular localization of tethers and their ability to interact with Rabs and soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) suggests that tethers participate in determining the specificity of membrane fusion. An accepted model of tether function considers them molecular “bridges” that link opposing membranes before SNARE pairing. This model has been extended by findings in various experimental systems, suggesting that tethers may have other functions. Recent reports implicate tethers in the assembly of SNARE complexes, cargo selection and transit, cytoskeletal events, and localized attachment of regulatory proteins. A concept of tethers as scaffolding machines that recruit protein components involved in varied cellular responses is emerging. In this model, tethers function as integration switches that simultaneously transmit information to coordinate distinct processes required for membrane traffic.

scholarly journals Syntaxin 6 Regulates Glut4 Trafficking in 3T3-L1 Adipocytes

2003 ◽  
Vol 14 (7) ◽  
pp. 2946-2958 ◽  
Author(s):  
H. Kumudu I. Perera ◽  
Mairi Clarke ◽  
Nicholas J. Morris ◽  
Wanjin Hong ◽  
Luke H. Chamberlain ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Trafficking ◽  
Glucose Transporter ◽  
Attachment Protein ◽  
Insulin Withdrawal ◽  
Sensitive Factor ◽  
Cytosolic Domain ◽  
Protein Receptors ◽  
Adipose Cells

Insulin stimulates the movement of glucose transporter-4 (Glut4)–containing vesicles to the plasma membrane of adipose cells. We investigated the role of post-Golgi t-soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) in the trafficking of Glut4 in 3T3-L1 adipocytes. Greater than 85% of syntaxin 6 was found in Glut4-containing vesicles, and this t-SNARE exhibited insulin-stimulated movement to the plasma membrane. In contrast, the colocalization of Glut4 with syntaxin 7, 8, or 12/13 was limited and these molecules did not translocate to the plasma membrane. We used adenovirus to overexpress the cytosolic domain of these syntaxin's and studied their effects on Glut4 traffic. Overexpression of the cytosolic domain of syntaxin 6 did not affect insulin-stimulated glucose transport, but increased basal deGlc transport and cell surface Glut4 levels. Moreover, the syntaxin 6 cytosolic domain significantly reduced the rate of Glut4 reinternalization after insulin withdrawal and perturbed subendosomal Glut4 sorting; the corresponding domains of syntaxins 8 and 12 were without effect. Our data suggest that syntaxin 6 is involved in a membrane-trafficking step that sequesters Glut4 away from traffic destined for the plasma membrane. We speculate that this is at the level of traffic of Glut4 into its unique storage compartment and that syntaxin 16 may be involved.

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