ArticleJournal of neuroscience methods2011
A novel method for producing mono-biotinylated, biologically active neurotrophic factors: an essential reagent for single molecule study of axonal transport.
Article in Journal of neuroscience methods, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 39 citations in OpenAlex.
- FRESH extrusion 3D printing of type-1 collagen hydrogels photocrosslinked using ruthenium.PloS one · 2025Article
- Failure to Thrive: Impaired BDNF Transport along the Cortical-Striatal Axis in Mouse Q140 Neurons of Huntington's Disease.Biology · 2023Article
- Age-induced nitrative stress decreases retrograde transport of proNGF via TrkA and increases proNGF retrograde transport and neurodegeneration via p75Frontiers in molecular neuroscience · 2023Article
- Retrograde Axonal Transport of Neurotrophins in Basal Forebrain Cholinergic Neurons.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Molecular-Scale Dynamics of Long Range Retrograde Brain-Derived Neurotrophic Factor Transport Shaped by Cellular Spatial Context.Frontiers in neuroscience · 2022Article
- Swedish Nerve Growth Factor Mutation (NGFThe Journal of neuroscience : the official journal of the Society for Neuroscience · 2018Article
- Inhibition of RhoA reduces propofol-mediated growth cone collapse, axonal transport impairment, loss of synaptic connectivity, and behavioural deficits.British journal of anaesthesia · 2018Article
- Aberrant GlyRS-HDAC6 interaction linked to axonal transport deficits in Charcot-Marie-Tooth neuropathy.Nature communications · 2018Article
- Article
- Synuclein impairs trafficking and signaling of BDNF in a mouse model of Parkinson's disease.Scientific reports · 2017Article
- An Optimized Procedure for the Site-Directed Labeling of NGF and proNGF for Imaging Purposes.Frontiers in molecular biosciences · 2017Article
- Article
- Amyloid precursor protein-mediated endocytic pathway disruption induces axonal dysfunction and neurodegeneration.The Journal of clinical investigation · 2016Article
- Ligand-induced dynamics of neurotrophin receptors investigated by single-molecule imaging approaches.International journal of molecular sciences · 2015Review
- Particle generation, functionalization and sortase A-mediated modification with targeting of single-chain antibodies for diagnostic and therapeutic use.Nature protocols · 2015Article
- Article
- Site-specific biotinylation of purified proteins using BirA.Methods in molecular biology (Clifton, N.J.) · 2015Article
- Real-time imaging of axonal transport of quantum dot-labeled BDNF in primary neurons.Journal of visualized experiments : JoVE · 2014Article
- Creation and Evaluation of a Single-chain Antibody Tetramer that Targets Brain Endothelial Cells.AIChE journal. American Institute of Chemical Engineers · 2014Article
- Site-specific labeling of neurotrophins and their receptors via short and versatile peptide tags.PloS one · 2014Article
Corrections and comments
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
Abstract
In this report, we describe a novel method for producing mature and biologically active mono-biotinylated nerve growth factors (mBtNGF) that can be used for single molecule studies of real-time movement of neurotrophins within axons of neurons. We inserted an AviTag sequence into the C-terminal of the full length mouse preproNGF cDNA and cloned the fusion construct into a pcDNA3.1 mammalian expression vector. We also subcloned the Escherichia coli biotin ligase, BirA, into a pcDNA3.1 vector. These two plasmids were then transiently co-expressed in HEK293FT cells. As a result, the AviTag located in the C-terminal of preproNGF was selectively ligated to a single biotin by BirA. The prepro sequence of NGF was subsequently cleaved within the cell. Mature mono-biotinylated NGF (mBtNGF) was secreted into cell culture media and was purified using Ni resin. We carried out activity assays and our results showed that mBtNGF retained biological activities that were comparable to normal NGF purified from mouse sub maxillary glands. We further verified the biotinylation efficiency of mBtNGF and the level of non-biotinylated NGF was virtually undetectable in the final preparation. Finally, by conjugating to quantum-dot streptavidin, mBtNGF was successfully used for single molecule study of axonal NGF trafficking in neurons.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.