Engineered botulinum neurotoxin B with improved efficacy for targeting human receptors
(2017) In Nature Communications 8(1). p.1-10- Abstract
Botulinum neurotoxin B is a Food and Drug Administration-approved therapeutic toxin. However, it has lower binding affinity toward the human version of its major receptor, synaptotagmin II (h-Syt II), compared to mouse Syt II, because of a residue difference. Increasing the binding affinity to h-Syt II may improve botulinum neurotoxin B's therapeutic efficacy and reduce adverse effects. Here we utilized the bacterial adenylate cyclase two-hybrid method and carried out a saturation mutagenesis screen in the Syt II-binding pocket of botulinum neurotoxin B. The screen identifies E1191 as a key residue: replacing it with M/C/V/Q enhances botulinum neurotoxin B binding to human synaptotagmin II. Adding S1199Y/W or W1178Q as a secondary... (More)
Botulinum neurotoxin B is a Food and Drug Administration-approved therapeutic toxin. However, it has lower binding affinity toward the human version of its major receptor, synaptotagmin II (h-Syt II), compared to mouse Syt II, because of a residue difference. Increasing the binding affinity to h-Syt II may improve botulinum neurotoxin B's therapeutic efficacy and reduce adverse effects. Here we utilized the bacterial adenylate cyclase two-hybrid method and carried out a saturation mutagenesis screen in the Syt II-binding pocket of botulinum neurotoxin B. The screen identifies E1191 as a key residue: replacing it with M/C/V/Q enhances botulinum neurotoxin B binding to human synaptotagmin II. Adding S1199Y/W or W1178Q as a secondary mutation further increases binding affinity. Mutant botulinum neurotoxin B containing E1191M/S1199Y exhibits ~11-fold higher efficacy in blocking neurotransmission than wild-type botulinum neurotoxin B in neurons expressing human synaptotagmin II, demonstrating that enhancing receptor binding increases the overall efficacy at functional levels. The engineered botulinum neurotoxin B provides a platform to develop therapeutic toxins with improved efficacy.Humans are less sensitive to the therapeutic effects of botulinum neurotoxin B (BoNT/B) than the animal models it is tested on due to differences between the human and the mouse receptors. Here, the authors engineer BoNT/B to improve its affinity to human receptors and enhance its therapeutic efficacy.
(Less)
- author
- Tao, Liang
; Peng, Lisheng
; Berntsson, Ronnie P-A
; Liu, Sai Man
; Park, SunHyun
; Yu, Feifan
; Boone, Christopher
; Palan, Shilpa
; Beard, Matthew
; Chabrier, Pierre-Etienne
; Stenmark, Pål
LU
; Krupp, Johannes
and Dong, Min
(Less) - publishing date
- 2017-07-03
- type
- Contribution to journal
- publication status
- published
- keywords
- Acetylcholine Release Inhibitors/pharmacology, Animals, Botulinum Toxins, Type A/genetics, Humans, Mutagenesis, Site-Directed, Neurons/drug effects, Patch-Clamp Techniques, Protein Binding/genetics, Rats, Recombinant Proteins, Synaptotagmin II/metabolism, Two-Hybrid System Techniques
- in
- Nature Communications
- volume
- 8
- issue
- 1
- pages
- 1 - 10
- publisher
- Nature Publishing Group
- external identifiers
-
- pmid:28674381
- scopus:85021734647
- ISSN
- 2041-1723
- DOI
- 10.1038/s41467-017-00064-y
- language
- English
- LU publication?
- no
- id
- edd20430-4997-4c57-bddb-2e87c97666e0
- date added to LUP
- 2019-04-30 07:55:03
- date last changed
- 2024-06-26 13:44:24
@article{edd20430-4997-4c57-bddb-2e87c97666e0,
abstract = {{<p>Botulinum neurotoxin B is a Food and Drug Administration-approved therapeutic toxin. However, it has lower binding affinity toward the human version of its major receptor, synaptotagmin II (h-Syt II), compared to mouse Syt II, because of a residue difference. Increasing the binding affinity to h-Syt II may improve botulinum neurotoxin B's therapeutic efficacy and reduce adverse effects. Here we utilized the bacterial adenylate cyclase two-hybrid method and carried out a saturation mutagenesis screen in the Syt II-binding pocket of botulinum neurotoxin B. The screen identifies E1191 as a key residue: replacing it with M/C/V/Q enhances botulinum neurotoxin B binding to human synaptotagmin II. Adding S1199Y/W or W1178Q as a secondary mutation further increases binding affinity. Mutant botulinum neurotoxin B containing E1191M/S1199Y exhibits ~11-fold higher efficacy in blocking neurotransmission than wild-type botulinum neurotoxin B in neurons expressing human synaptotagmin II, demonstrating that enhancing receptor binding increases the overall efficacy at functional levels. The engineered botulinum neurotoxin B provides a platform to develop therapeutic toxins with improved efficacy.Humans are less sensitive to the therapeutic effects of botulinum neurotoxin B (BoNT/B) than the animal models it is tested on due to differences between the human and the mouse receptors. Here, the authors engineer BoNT/B to improve its affinity to human receptors and enhance its therapeutic efficacy.</p>}},
author = {{Tao, Liang and Peng, Lisheng and Berntsson, Ronnie P-A and Liu, Sai Man and Park, SunHyun and Yu, Feifan and Boone, Christopher and Palan, Shilpa and Beard, Matthew and Chabrier, Pierre-Etienne and Stenmark, Pål and Krupp, Johannes and Dong, Min}},
issn = {{2041-1723}},
keywords = {{Acetylcholine Release Inhibitors/pharmacology; Animals; Botulinum Toxins, Type A/genetics; Humans; Mutagenesis, Site-Directed; Neurons/drug effects; Patch-Clamp Techniques; Protein Binding/genetics; Rats; Recombinant Proteins; Synaptotagmin II/metabolism; Two-Hybrid System Techniques}},
language = {{eng}},
month = {{07}},
number = {{1}},
pages = {{1--10}},
publisher = {{Nature Publishing Group}},
series = {{Nature Communications}},
title = {{Engineered botulinum neurotoxin B with improved efficacy for targeting human receptors}},
url = {{http://dx.doi.org/10.1038/s41467-017-00064-y}},
doi = {{10.1038/s41467-017-00064-y}},
volume = {{8}},
year = {{2017}},
}