title: Stapling of the botulinum type A protease to growth factors and neuropeptides allows selective targeting of neuroendocrine cells.
abstract: Precise cellular targeting of macromolecular cargos has important biotechnological and medical implications. Using a recently established 'protein stapling' method, we linked the proteolytic domain of botulinum neurotoxin type A (BoNT/A) to a selection of ligands to target neuroendocrine tumor cells. The botulinum proteolytic domain was chosen because of its well-known potency to block the release of neurotransmitters and hormones. Among nine tested stapled ligands, the epidermal growth factor was able to deliver the botulinum enzyme into pheochromocytoma PC12 and insulinoma Min6 cells; ciliary neurotrophic factor was effective on neuroblastoma SH-SY5Y and Neuro2A cells, whereas corticotropin-releasing hormone was active on pituitary AtT-20 cells and the two neuroblastoma cell lines. In neuronal cultures, the epidermal growth factor- and ciliary neurotrophic factor-directed botulinum enzyme targeted distinct subsets of neurons whereas the whole native neurotoxin targeted the cortical neurons indiscriminately. At nanomolar concentrations, the retargeted botulinum molecules were able to inhibit stimulated release of hormones from tested cell lines suggesting their application for treatments of neuroendocrine disorders.
authors: MRC Laboratory of Molecular Biology, Neurobiology, Cambridge, UK.
authors: Jason
authors: J
authors: Arsenault
authors: Enrico
authors: E
authors: Ferrari
authors: Dhevahi
authors: D
authors: Niranjan
authors: Sabine A G
authors: SA
authors: Cuijpers
authors: Chunjing
authors: C
authors: Gu
authors: Yvonne
authors: Y
authors: Vallis
authors: John
authors: J
authors: O'Brien
authors: Bazbek
authors: B
authors: Davletov
mesh_terms: D000818:Animals; D019274:Botulinum Toxins, Type A / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002460:Cell Line; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009447:Neuroblastoma / Q000473:pathology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D009479:Neuropeptides / Q000737:chemistry / Q000378:metabolism*; D009638:Norepinephrine / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D017434:Protein Structure, Tertiary / Q000187:drug effects; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism; D014316:Tritium / Q000378:metabolism
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins; D009479:Neuropeptides; D011993:Recombinant Fusion Proteins; D050825:Synaptosomal-Associated Protein 25; D014316:Tritium; D011189:Potassium Chloride; D019274:Botulinum Toxins, Type A; D009638:Norepinephrine
keywords: SNAP25; botulinum; growth factor; neuroendocrine tumor; neuropeptide; targeting
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