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Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation
The spider venom protein, double-knot toxin (DkTx), partitions into the cellular membrane and binds bivalently to the pain-sensing ion channel, TRPV1, triggering long-lasting channel activation. In contrast, its monovalent single knots membrane partition poorly and invoke rapidly reversible TRPV1 ac...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10404664/ https://www.ncbi.nlm.nih.gov/pubmed/37302551 http://dx.doi.org/10.1016/j.jbc.2023.104903 |
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author | Singh, Yashaswi Sarkar, Debayan Duari, Subhadeep G, Shashaank Indra Guru, Pawas Kumar M V, Hrishikesh Singh, Dheerendra Bhardwaj, Sahil Kalia, Jeet |
author_facet | Singh, Yashaswi Sarkar, Debayan Duari, Subhadeep G, Shashaank Indra Guru, Pawas Kumar M V, Hrishikesh Singh, Dheerendra Bhardwaj, Sahil Kalia, Jeet |
author_sort | Singh, Yashaswi |
collection | PubMed |
description | The spider venom protein, double-knot toxin (DkTx), partitions into the cellular membrane and binds bivalently to the pain-sensing ion channel, TRPV1, triggering long-lasting channel activation. In contrast, its monovalent single knots membrane partition poorly and invoke rapidly reversible TRPV1 activation. To discern the contributions of the bivalency and membrane affinity of DkTx to its sustained mode of action, here, we developed diverse toxin variants including those containing truncated linkers between individual knots, precluding bivalent binding. Additionally, by appending the single-knot domains to the Kv2.1 channel-targeting toxin, SGTx, we created monovalent double-knot proteins that demonstrated higher membrane affinity and more sustained TRPV1 activation than the single-knots. We also produced hyper-membrane affinity-possessing tetra-knot proteins, (DkTx)(2) and DkTx-(SGTx)(2), that demonstrated longer-lasting TRPV1 activation than DkTx, establishing the central role of the membrane affinity of DkTx in endowing it with its sustained TRPV1 activation properties. These results suggest that high membrane affinity-possessing TRPV1 agonists can potentially serve as long-acting analgesics. |
format | Online Article Text |
id | pubmed-10404664 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-104046642023-08-08 Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation Singh, Yashaswi Sarkar, Debayan Duari, Subhadeep G, Shashaank Indra Guru, Pawas Kumar M V, Hrishikesh Singh, Dheerendra Bhardwaj, Sahil Kalia, Jeet J Biol Chem Research Article The spider venom protein, double-knot toxin (DkTx), partitions into the cellular membrane and binds bivalently to the pain-sensing ion channel, TRPV1, triggering long-lasting channel activation. In contrast, its monovalent single knots membrane partition poorly and invoke rapidly reversible TRPV1 activation. To discern the contributions of the bivalency and membrane affinity of DkTx to its sustained mode of action, here, we developed diverse toxin variants including those containing truncated linkers between individual knots, precluding bivalent binding. Additionally, by appending the single-knot domains to the Kv2.1 channel-targeting toxin, SGTx, we created monovalent double-knot proteins that demonstrated higher membrane affinity and more sustained TRPV1 activation than the single-knots. We also produced hyper-membrane affinity-possessing tetra-knot proteins, (DkTx)(2) and DkTx-(SGTx)(2), that demonstrated longer-lasting TRPV1 activation than DkTx, establishing the central role of the membrane affinity of DkTx in endowing it with its sustained TRPV1 activation properties. These results suggest that high membrane affinity-possessing TRPV1 agonists can potentially serve as long-acting analgesics. American Society for Biochemistry and Molecular Biology 2023-06-10 /pmc/articles/PMC10404664/ /pubmed/37302551 http://dx.doi.org/10.1016/j.jbc.2023.104903 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Singh, Yashaswi Sarkar, Debayan Duari, Subhadeep G, Shashaank Indra Guru, Pawas Kumar M V, Hrishikesh Singh, Dheerendra Bhardwaj, Sahil Kalia, Jeet Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation |
title | Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation |
title_full | Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation |
title_fullStr | Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation |
title_full_unstemmed | Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation |
title_short | Dissecting the contributions of membrane affinity and bivalency of the spider venom protein DkTx to its sustained mode of TRPV1 activation |
title_sort | dissecting the contributions of membrane affinity and bivalency of the spider venom protein dktx to its sustained mode of trpv1 activation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10404664/ https://www.ncbi.nlm.nih.gov/pubmed/37302551 http://dx.doi.org/10.1016/j.jbc.2023.104903 |
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