ArticleProceedings of the National Academy of Sciences of the United States of America2006
Construction of a mutated pro-nerve growth factor resistant to degradation and suitable for biophysical and cellular utilization.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- Unraveling proteomic signatures and neuroinflammatory networks in a CCI rat model of early sciatica: insights for neuropathic pain mechanisms.Frontiers in molecular neuroscience · 2025Article
- The expression system affects the binding affinity between p75NTR and proNGF.Biochemistry and biophysics reports · 2024Article
- Accelerated neurodegeneration of basal forebrain cholinergic neurons in HIV-1 gp120 transgenic mice: Critical role of the p75 neurotrophin receptor.Brain, behavior, and immunity · 2024Article
- Pro region engineering of nerve growth factor by deep mutational scanning enables a yeast platform for conformational epitope mapping of anti-NGF monoclonal antibodies.Biotechnology and bioengineering · 2018Article
- Inducible overexpression of endothelial proNGF as a mouse model to study microvascular dysfunction.Biochimica et biophysica acta. Molecular basis of disease · 2018Article
- Conformational characterization of nerve growth factor-β reveals that its regulatory pro-part domain stabilizes three loop regions in its mature part.The Journal of biological chemistry · 2017Article
- ProNGF, but Not NGF, Switches from Neurotrophic to Apoptotic Activity in Response to Reductions in TrkA Receptor Levels.International journal of molecular sciences · 2017Article
- A Pro-Nerve Growth Factor (proNGF) and NGF Binding Protein, α2-Macroglobulin, Differentially Regulates p75 and TrkA Receptors and Is Relevant to Neurodegeneration Ex Vivo and In Vivo.Molecular and cellular biology · 2015Article
- Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR).International journal of biochemistry and molecular biology · 2015Article
- ProNGF derived from rat sciatic nerves downregulates neurite elongation and axon specification in PC12 cells.Frontiers in cellular neuroscience · 2015Article
- The p75NTR signaling cascade mediates mechanical hyperalgesia induced by nerve growth factor injected into the rat hind paw.Neuroscience · 2013Article
- Modulation of p75(NTR) prevents diabetes- and proNGF-induced retinal inflammation and blood-retina barrier breakdown in mice and rats.Diabetologia · 2013Article
- ProNGF promotes neurite growth from a subset of NGF-dependent neurons by a p75NTR-dependent mechanism.Development (Cambridge, England) · 2013Article
- Cardiac fibroblasts regulate sympathetic nerve sprouting and neurocardiac synapse stability.PloS one · 2013Article
- Pro-nerve growth factor induces autocrine stimulation of breast cancer cell invasion through tropomyosin-related kinase A (TrkA) and sortilin protein.The Journal of biological chemistry · 2012Article
- Electroporation-mediated gene delivery of cleavage-resistant pro-nerve growth factor causes retinal neuro- and vascular degeneration.Molecular vision · 2012Article
- Human tryptase cleaves pro-nerve growth factor (pro-NGF): hints of local, mast cell-dependent regulation of NGF/pro-NGF action.The Journal of biological chemistry · 2011Article
- Alteration of growth factors and neuronal death in diabetic retinopathy: what we have learned so far.Molecular vision · 2011Review
- Conformational plasticity of proNGF.PloS one · 2011Article
- ProNGF mediates death of Natural Killer cells through activation of the p75NTR-sortilin complex.Journal of neuroimmunology · 2010Article
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Authors and funding
3 authors.
Funding
Abstract
Precursor of nerve growth factor (proNGF) has been found to be proapoptotic in several cell types and mediates its effects by binding to p75 neurotrophin receptor (p75NTR) and sortilin. The proNGF molecule is processed by proteases at three dibasic sites found in the pro domain to form mature NGF (termed herein as sites 1, 2, and 3 from the proNGF N terminus). Of these processing sites, site 3, adjacent to the N terminus of mature NGF, was thought to be the major site responsible for processing of proNGF to mature NGF. We found that mutating this major processing site (site 3) resulted in a form of proNGF that was only partially stable. On introducing additional mutations in the pro domain at the other two dibasic sites, we found the stability of proNGF to increase significantly. Here we describe the construction, expression, and purification of this more stable proNGF molecule. The two consecutive basic residues at each of the three sites were mutated to neutral alanine residues. Expression was performed in stably transfected Sf21 insect cells. Purification involved strong cation-exchange chromatography and N60 immunoaffinity column chromatography. The construct with all three sites mutated (termed proNGF123) gave all proNGF with no mature NGF and was not cleaved by three proconvertases (furin, PACE-4, and PC-2) known to proteolyze proneurotrophins in vivo. This stable proNGF molecule demonstrated proapoptotic activity on rat pheocytochroma PC12 cells, PC12nnr cells, C6 glioblastoma cells, and RN22 schwannoma cells.
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