HBpF-proBDNF: A New Tool for the Analysis of Pro-Brain Derived Neurotrophic Factor Receptor Signaling and Cell Biology
| Autor: | Andrés de Léon, Vincent Soubannier, Philip A. Barker, Julien Gibon, Geneviève Dorval, Perrine Gaub, Philippe Séguéla |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Male B Vitamins Receptor complex lcsh:Medicine Apoptosis Tropomyosin receptor kinase B PC12 Cells Biochemistry Mice 0302 clinical medicine Neurotrophic factors Animal Cells Protein Isoforms Enzyme Inhibitors lcsh:Science Neurons Multidisciplinary biology Cell Death Organic Compounds Signal transducing adaptor protein Proteases Vitamins Cell biology Enzymes Chemistry Cell Processes Physical Sciences Signal transduction Cellular Types Neurotrophin Signal Transduction Research Article Cytological Techniques Immunoblotting Biotin Molecular Probe Techniques Nerve Tissue Proteins Receptors Nerve Growth Factor Research and Analysis Methods 03 medical and health sciences Affinity Purification Animals Humans Protease Inhibitors Protein Precursors Molecular Biology Techniques Molecular Biology Imidazole Brain-derived neurotrophic factor Brain-Derived Neurotrophic Factor HEK 293 cells Organic Chemistry lcsh:R Chemical Compounds Biology and Life Sciences Proteins Cell Biology Rats Adaptor Proteins Vesicular Transport 030104 developmental biology HEK293 Cells biology.protein Enzymology lcsh:Q 030217 neurology & neurosurgery Purification Techniques |
| Zdroj: | PLoS ONE, Vol 11, Iss 3, p e0150601 (2016) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Neurotrophins activate intracellular signaling pathways necessary for neuronal survival, growth and apoptosis. The most abundant neurotrophin in the adult brain, brain-derived neurotrophic factor (BDNF), is first synthesized as a proBDNF precursor and recent studies have demonstrated that proBDNF can be secreted and that it functions as a ligand for a receptor complex containing p75NTR and sortilin. Activation of proBDNF receptors mediates growth cone collapse, reduces synaptic activity, and facilitates developmental apoptosis of motoneurons but the precise signaling cascades have been difficult to discern. To address this, we have engineered, expressed and purified HBpF-proBDNF, an expression construct containing a 6X-HIS tag, a biotin acceptor peptide (BAP) sequence, a PreScission™ Protease cleavage site and a FLAG-tag attached to the N-terminal part of murine proBDNF. Intact HBpF-proBDNF has activities indistinguishable from its wild-type counterpart and can be used to purify proBDNF signaling complexes or to monitor proBDNF endocytosis and retrograde transport. HBpF-proBDNF will be useful for characterizing proBDNF signaling complexes and for deciphering the role of proBDNF in neuronal development, synapse function and neurodegenerative disease. |
| Databáze: | OpenAIRE |
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