Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model
Abstract Purpose: RNYK is a selective agonist of the neurotrophic tyrosine kinase receptor type 2 (NTRK2) which has been screened from a phage-displayed peptide library. Its sequence is SGVYKVAYDWQH, similar to a native NTRK2 ligand, that is, brain-derived neurotrophic factor (BDNF). The current stu...
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| Format: | Article |
| Language: | English |
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Knowledge E
2024-03-01
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| Series: | Journal of Ophthalmic & Vision Research |
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| Online Access: | https://doi.org/10.18502/jovr.v19i1.15439 |
| _version_ | 1797230386081693696 |
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| author | Fatemeh Nafian Shahin Yazdani Mohammad Javad Rasaee Babak Kamali Doust Azad Narsis Daftarian Mozhgan Rezaei Kanavi |
| author_facet | Fatemeh Nafian Shahin Yazdani Mohammad Javad Rasaee Babak Kamali Doust Azad Narsis Daftarian Mozhgan Rezaei Kanavi |
| author_sort | Fatemeh Nafian |
| collection | DOAJ |
| description | Abstract Purpose: RNYK is a selective agonist of the neurotrophic tyrosine kinase receptor type 2 (NTRK2) which has been screened from a phage-displayed peptide library. Its sequence is SGVYKVAYDWQH, similar to a native NTRK2 ligand, that is, brain-derived neurotrophic factor (BDNF). The current study was performed to recognize and confirm critical residues for RNYK activity in a glaucoma-on-a-chip model. Methods: We designed a modified RNYK (mRNYK) peptide based on hotspots of the RNYK sequence identified by alanine scanning. The critical residues consisted of tyrosine, valine, aspartic acid, and tryptophan (YVDW); however, lysine and glutamine were also maintained in the final sequence (YKVDWQ) for forming amide bonds and peptide dimerization. The affinity of mRNYK binding was confirmed by testing against NTRK2 receptors on the surface of ATRA-treated SH-SY5Y cells. The neuroprotective effect of mRNYK was also evaluated in cell culture after elevated pressure insult in a glaucoma-on-a-chip model. Results: The primary amine on the lysine side-chain from one sequence (YKVDWQ) reacted with a γ-carboxamide group of glutamine from the other sequence, forming dimeric mRNYK. In silico, molecular dynamic simulations of the mRNYK–NTRK2 complex showed more stable and stronger interactions as compared to the RNYK–NTRK2 complex. In vitro, mRNYK demonstrated a neuroprotective effect on SH-SY5Y cells under normal and elevated pressure comparable to RNYK. The 50% effective concentration (logEC50) for mRNYK was 0.7009, which was better than RNYK with a logEC50 of 0.8318. Conclusion: The modified peptide studied herein showed improved stability over the original peptide (RNYK) and demonstrated potential for use as a BDNF agonist with neuroprotective properties for treatment of neurodegenerative disorders such as glaucoma. |
| first_indexed | 2024-04-24T15:27:39Z |
| format | Article |
| id | doaj.art-59e995f92eed44ccad282a3b8e54c41c |
| institution | Directory Open Access Journal |
| issn | 2008-2010 2008-322X |
| language | English |
| last_indexed | 2024-04-24T15:27:39Z |
| publishDate | 2024-03-01 |
| publisher | Knowledge E |
| record_format | Article |
| series | Journal of Ophthalmic & Vision Research |
| spelling | doaj.art-59e995f92eed44ccad282a3b8e54c41c2024-04-02T05:30:46ZengKnowledge EJournal of Ophthalmic & Vision Research2008-20102008-322X2024-03-01191587010.18502/jovr.v19i1.15439jovr.v19i1.15439Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip ModelFatemeh Nafian0Shahin Yazdani1Mohammad Javad Rasaee2Babak Kamali Doust Azad3Narsis Daftarian4Mozhgan Rezaei Kanavi5 Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran School of Electrical and Computer Engineering, Tehran University, Tehran, Iran Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran.Abstract Purpose: RNYK is a selective agonist of the neurotrophic tyrosine kinase receptor type 2 (NTRK2) which has been screened from a phage-displayed peptide library. Its sequence is SGVYKVAYDWQH, similar to a native NTRK2 ligand, that is, brain-derived neurotrophic factor (BDNF). The current study was performed to recognize and confirm critical residues for RNYK activity in a glaucoma-on-a-chip model. Methods: We designed a modified RNYK (mRNYK) peptide based on hotspots of the RNYK sequence identified by alanine scanning. The critical residues consisted of tyrosine, valine, aspartic acid, and tryptophan (YVDW); however, lysine and glutamine were also maintained in the final sequence (YKVDWQ) for forming amide bonds and peptide dimerization. The affinity of mRNYK binding was confirmed by testing against NTRK2 receptors on the surface of ATRA-treated SH-SY5Y cells. The neuroprotective effect of mRNYK was also evaluated in cell culture after elevated pressure insult in a glaucoma-on-a-chip model. Results: The primary amine on the lysine side-chain from one sequence (YKVDWQ) reacted with a γ-carboxamide group of glutamine from the other sequence, forming dimeric mRNYK. In silico, molecular dynamic simulations of the mRNYK–NTRK2 complex showed more stable and stronger interactions as compared to the RNYK–NTRK2 complex. In vitro, mRNYK demonstrated a neuroprotective effect on SH-SY5Y cells under normal and elevated pressure comparable to RNYK. The 50% effective concentration (logEC50) for mRNYK was 0.7009, which was better than RNYK with a logEC50 of 0.8318. Conclusion: The modified peptide studied herein showed improved stability over the original peptide (RNYK) and demonstrated potential for use as a BDNF agonist with neuroprotective properties for treatment of neurodegenerative disorders such as glaucoma.https://doi.org/10.18502/jovr.v19i1.15439agonist brain-derived neurotrophic factor neuroprotection neurotrophic tyrosine receptor kinase |
| spellingShingle | Fatemeh Nafian Shahin Yazdani Mohammad Javad Rasaee Babak Kamali Doust Azad Narsis Daftarian Mozhgan Rezaei Kanavi Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model Journal of Ophthalmic & Vision Research agonist brain-derived neurotrophic factor neuroprotection neurotrophic tyrosine receptor kinase |
| title | Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model |
| title_full | Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model |
| title_fullStr | Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model |
| title_full_unstemmed | Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model |
| title_short | Modification of a Selective NTRK2 Agonist and Confirmation of Activity in a Glaucoma-on-a-Chip Model |
| title_sort | modification of a selective ntrk2 agonist and confirmation of activity in a glaucoma on a chip model |
| topic | agonist brain-derived neurotrophic factor neuroprotection neurotrophic tyrosine receptor kinase |
| url | https://doi.org/10.18502/jovr.v19i1.15439 |
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