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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...

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Autores principales: Singh, Yashaswi, Sarkar, Debayan, Duari, Subhadeep, G, Shashaank, Indra Guru, Pawas Kumar, M V, Hrishikesh, Singh, Dheerendra, Bhardwaj, Sahil, Kalia, Jeet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2023
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.
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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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