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Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers
It is widely accepted that snake venom cardiotoxins (CTXs) target the plasma membranes of cells. In the present study, we investigated the role of Asp residues in the interaction of Naja atra cardiotoxin 1 (CTX1) and cardiotoxin 3 (CTX3) with phospholipid bilayers using chemical modification. CTX1 c...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7232319/ https://www.ncbi.nlm.nih.gov/pubmed/32325789 http://dx.doi.org/10.3390/toxins12040262 |
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author | Wu, Guan-Lin Shi, Yi-Jun Huang, Chia-Hui Lee, Yuan-Chin Wang, Liang-Jun Chiou, Jing-Ting Lu, Chi-Yu Chang, Long-Sen |
author_facet | Wu, Guan-Lin Shi, Yi-Jun Huang, Chia-Hui Lee, Yuan-Chin Wang, Liang-Jun Chiou, Jing-Ting Lu, Chi-Yu Chang, Long-Sen |
author_sort | Wu, Guan-Lin |
collection | PubMed |
description | It is widely accepted that snake venom cardiotoxins (CTXs) target the plasma membranes of cells. In the present study, we investigated the role of Asp residues in the interaction of Naja atra cardiotoxin 1 (CTX1) and cardiotoxin 3 (CTX3) with phospholipid bilayers using chemical modification. CTX1 contains three Asp residues at positions 29, 40, and 57; CTX3 contains two Asp residues at positions 40 and 57. Compared to Asp29 and Asp40, Asp57 was sparingly modified with semi-carbazide, as revealed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass and mass/mass analyses. Thus, semi-carbazide-modified CTX1 (SEM-CTX1) mainly contained modified Asp29 and Asp40, while SEM-CTX3 contained modified Asp40. Compared to that of native toxins, trifluoroethanol easily induced structural transition of SEM-CTX1 and SEM-CTX3, suggesting that the structural flexibility of CTXs was constrained by Asp40. Modification of Asp29 and Asp40 markedly promoted the ability of CTX1 to induce permeability of cell membranes and lipid vesicles; CTX3 and SEM-CTX3 showed similar membrane-damaging activity. Modification of Asp residues did not affect the membrane-binding capability of CTXs. Circular dichroism spectra of SEM-CTX3 and CTX3 were similar, while the gross conformation of SEM-CTX1 was distinct from that of CTX1. The interaction of CTX1 with membrane was distinctly changed by Asp modification. Collectively, our data suggest that Asp29 of CTX1 suppresses the optimization of membrane-bound conformation to a fully active state and that the function of Asp40 in the structural constraints of CTX1 and CTX3 is not important for the manifestation of membrane-perturbing activity. |
format | Online Article Text |
id | pubmed-7232319 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-72323192020-05-22 Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers Wu, Guan-Lin Shi, Yi-Jun Huang, Chia-Hui Lee, Yuan-Chin Wang, Liang-Jun Chiou, Jing-Ting Lu, Chi-Yu Chang, Long-Sen Toxins (Basel) Article It is widely accepted that snake venom cardiotoxins (CTXs) target the plasma membranes of cells. In the present study, we investigated the role of Asp residues in the interaction of Naja atra cardiotoxin 1 (CTX1) and cardiotoxin 3 (CTX3) with phospholipid bilayers using chemical modification. CTX1 contains three Asp residues at positions 29, 40, and 57; CTX3 contains two Asp residues at positions 40 and 57. Compared to Asp29 and Asp40, Asp57 was sparingly modified with semi-carbazide, as revealed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass and mass/mass analyses. Thus, semi-carbazide-modified CTX1 (SEM-CTX1) mainly contained modified Asp29 and Asp40, while SEM-CTX3 contained modified Asp40. Compared to that of native toxins, trifluoroethanol easily induced structural transition of SEM-CTX1 and SEM-CTX3, suggesting that the structural flexibility of CTXs was constrained by Asp40. Modification of Asp29 and Asp40 markedly promoted the ability of CTX1 to induce permeability of cell membranes and lipid vesicles; CTX3 and SEM-CTX3 showed similar membrane-damaging activity. Modification of Asp residues did not affect the membrane-binding capability of CTXs. Circular dichroism spectra of SEM-CTX3 and CTX3 were similar, while the gross conformation of SEM-CTX1 was distinct from that of CTX1. The interaction of CTX1 with membrane was distinctly changed by Asp modification. Collectively, our data suggest that Asp29 of CTX1 suppresses the optimization of membrane-bound conformation to a fully active state and that the function of Asp40 in the structural constraints of CTX1 and CTX3 is not important for the manifestation of membrane-perturbing activity. MDPI 2020-04-18 /pmc/articles/PMC7232319/ /pubmed/32325789 http://dx.doi.org/10.3390/toxins12040262 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Wu, Guan-Lin Shi, Yi-Jun Huang, Chia-Hui Lee, Yuan-Chin Wang, Liang-Jun Chiou, Jing-Ting Lu, Chi-Yu Chang, Long-Sen Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers |
title | Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers |
title_full | Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers |
title_fullStr | Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers |
title_full_unstemmed | Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers |
title_short | Status of Asp29 and Asp40 in the Interaction of Naja atra Cardiotoxins with Lipid Bilayers |
title_sort | status of asp29 and asp40 in the interaction of naja atra cardiotoxins with lipid bilayers |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7232319/ https://www.ncbi.nlm.nih.gov/pubmed/32325789 http://dx.doi.org/10.3390/toxins12040262 |
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