title: HBpF-proBDNF: A New Tool for the Analysis of Pro-Brain Derived Neurotrophic Factor Receptor Signaling and Cell Biology.
abstract: 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.
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Perrine
authors.initials: P
authors.lastname: Gaub
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Andrès
authors.initials: A
authors.lastname: de Léon
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Julien
authors.initials: J
authors.lastname: Gibon
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Vincent
authors.initials: V
authors.lastname: Soubannier
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Dorval
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Philippe
authors.initials: P
authors.lastname: Séguéla
authors.affiliation: Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Philip A
authors.initials: PA
authors.lastname: Barker
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003584:Cytological Techniques / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D016716:PC12 Cells; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism; D015398:Signal Transduction; D000099183:Sortilin
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D019208:Brain-Derived Neurotrophic Factor; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D011498:Protein Precursors; D017475:Receptors, Nerve Growth Factor; D000099183:Sortilin; C484065:NGFR protein, human