Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry
Background : Numerous spider toxins are of interest as tools for neurophysiological research or as lead molecules for the development of pharmaceuticals and insecticides. Direct detection and identification of the interacting proteins of a spider toxin are helpful for its action-mechanism analysis a...
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ftdoajarticles:oai:doaj.org/article:33c065fd0b8c43b5940183fc9ac009a7 2023-05-15T15:08:12+02:00 Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry Hai Yu Hui Liu Yizhong Yan Zhigui Duan Xianchun Wang 2014-05-01T00:00:00Z https://doi.org/10.1186/1678-9199-20-18 https://doaj.org/article/33c065fd0b8c43b5940183fc9ac009a7 EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992014000200328&lng=en&tlng=en https://doaj.org/toc/1678-9199 1678-9199 doi:10.1186/1678-9199-20-18 https://doaj.org/article/33c065fd0b8c43b5940183fc9ac009a7 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 20, Iss 0 (2014) Biotin labeling Huwentoxin-IV Far-western blotting Interacting protein Mass spectrometry Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2014 ftdoajarticles https://doi.org/10.1186/1678-9199-20-18 2022-12-31T11:56:32Z Background : Numerous spider toxins are of interest as tools for neurophysiological research or as lead molecules for the development of pharmaceuticals and insecticides. Direct detection and identification of the interacting proteins of a spider toxin are helpful for its action-mechanism analysis and practical application. The present study employed a combinative strategy for the analysis of interacting proteins of huwentoxin-IV (HWTX-IV), a peptidic neurotoxin from the venom of the spiderSelenocosmia huwena.Results : HWTX-IV was first lightly labeled with biotin under the optimized mild experimental conditions and the toxin labeled with a single biotin group (monobiotinylated HWTX-IV) was demonstrated by electrophysiological experiments to retain its original bioactivity and was used in combination with far-western blotting to detect its interacting proteins. Comparative experiments indicated that some membrane proteins from rat neuromuscular junction preparations bind to monobiotinylated HWTX-IV after being transferred onto a PVDF membrane from the SDS-gel. With capillary high performance liquid chromatography-tandem mass spectrometry, several membrane proteins with which HWTX-IV potentially interacted were identified from the preparations and then bioinformatically analyzed.Conclusions : This work has provided not only a new insight into the action mechanism of HWTX-IV but also a reference technology for the relevant researches. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Journal of Venomous Animals and Toxins including Tropical Diseases 20 1 18 |
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Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Biotin labeling Huwentoxin-IV Far-western blotting Interacting protein Mass spectrometry Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
spellingShingle |
Biotin labeling Huwentoxin-IV Far-western blotting Interacting protein Mass spectrometry Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Hai Yu Hui Liu Yizhong Yan Zhigui Duan Xianchun Wang Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
topic_facet |
Biotin labeling Huwentoxin-IV Far-western blotting Interacting protein Mass spectrometry Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
Background : Numerous spider toxins are of interest as tools for neurophysiological research or as lead molecules for the development of pharmaceuticals and insecticides. Direct detection and identification of the interacting proteins of a spider toxin are helpful for its action-mechanism analysis and practical application. The present study employed a combinative strategy for the analysis of interacting proteins of huwentoxin-IV (HWTX-IV), a peptidic neurotoxin from the venom of the spiderSelenocosmia huwena.Results : HWTX-IV was first lightly labeled with biotin under the optimized mild experimental conditions and the toxin labeled with a single biotin group (monobiotinylated HWTX-IV) was demonstrated by electrophysiological experiments to retain its original bioactivity and was used in combination with far-western blotting to detect its interacting proteins. Comparative experiments indicated that some membrane proteins from rat neuromuscular junction preparations bind to monobiotinylated HWTX-IV after being transferred onto a PVDF membrane from the SDS-gel. With capillary high performance liquid chromatography-tandem mass spectrometry, several membrane proteins with which HWTX-IV potentially interacted were identified from the preparations and then bioinformatically analyzed.Conclusions : This work has provided not only a new insight into the action mechanism of HWTX-IV but also a reference technology for the relevant researches. |
format |
Article in Journal/Newspaper |
author |
Hai Yu Hui Liu Yizhong Yan Zhigui Duan Xianchun Wang |
author_facet |
Hai Yu Hui Liu Yizhong Yan Zhigui Duan Xianchun Wang |
author_sort |
Hai Yu |
title |
Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
title_short |
Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
title_full |
Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
title_fullStr |
Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
title_full_unstemmed |
Detection and identification of huwentoxin-IV interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
title_sort |
detection and identification of huwentoxin-iv interacting proteins by biotin-avidin chemistry combined with mass spectrometry |
publisher |
SciELO |
publishDate |
2014 |
url |
https://doi.org/10.1186/1678-9199-20-18 https://doaj.org/article/33c065fd0b8c43b5940183fc9ac009a7 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 20, Iss 0 (2014) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992014000200328&lng=en&tlng=en https://doaj.org/toc/1678-9199 1678-9199 doi:10.1186/1678-9199-20-18 https://doaj.org/article/33c065fd0b8c43b5940183fc9ac009a7 |
op_doi |
https://doi.org/10.1186/1678-9199-20-18 |
container_title |
Journal of Venomous Animals and Toxins including Tropical Diseases |
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20 |
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18 |
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1766339616720289792 |