The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes

ABSTRACT Botulinum neurotoxin (BoNT) is the causative agent of botulism and a widely used pharmaceutical to treat a variety of neurological diseases. BoNTs are 150-kDa protein toxins organized into heavy chain (HC) and light chain (LC) domains linked by a disulfide bond. The HC selectively binds to...

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Autores principales:	Sabine Pellett, Marite Bradshaw, William H. Tepp, Christina L. Pier, Regina C. M. Whitemarsh, Chen Chen, Joseph T. Barbieri, Eric A. Johnson
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	BoNT botulinum neurotoxin duration of action hybrid toxin subtype Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/aedc52e97f8748b7906c9ad8d9dcb24e
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spelling	oai:doaj.org-article:aedc52e97f8748b7906c9ad8d9dcb24e2021-11-15T15:53:26ZThe Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes10.1128/mBio.00089-182150-7511https://doaj.org/article/aedc52e97f8748b7906c9ad8d9dcb24e2018-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00089-18https://doaj.org/toc/2150-7511ABSTRACT Botulinum neurotoxin (BoNT) is the causative agent of botulism and a widely used pharmaceutical to treat a variety of neurological diseases. BoNTs are 150-kDa protein toxins organized into heavy chain (HC) and light chain (LC) domains linked by a disulfide bond. The HC selectively binds to neurons and aids cell entry of the enzymatically active LC. There are seven immunological BoNT serotypes (A to G); each serotype includes genetic variants, termed subtypes. Only two subtypes, BoNT/A1 and BoNT/B1, are currently used as therapeutics. BoNT serotype A (BoNT/A) subtypes A2 to A8 show distinct potency, duration of action, and pathology relative to BoNT/A1. Specifically, BoNT/A3 possesses shorter duration of action and elicits distinct symptoms in mice at high toxin doses. In this report, we analyzed the roles of LC and HC of BoNT/A3 for duration of action, neuronal cell entry, and mouse pathology by using clostridium-derived recombinant hybrid BoNTs consisting of reciprocal LC and HC (BoNTA1/A3 and BoNTA3/A1). Hybrid toxins were processed in their expression host to a dichain BoNT consisting of LC and HC linked via a disulfide bond. The LC and HC defined BoNT potency in mice and BoNT toxicity for cultured neuronal cells, while the LC defined the duration of BoNT action in cell and mouse models. Protein alignment identified a previously unrecognized region within the LC subtype A3 (LC/A3) relative to the other LC serotype A (LC/A) subtypes (low primary acid homology [LPH]) that correlated to intracellular LC localization. This study shows the utility of recombinant hybrid BoNTs with new therapeutic potential, while remaining sensitive to antitoxins and therapies to native BoNT. IMPORTANCE Botulinum neurotoxins are the most potent protein toxins for humans and potential bioterrorism threats, but they are also widely used as pharmaceuticals. Within the large family of BoNTs, only two subtypes are currently used as pharmaceuticals, with a large number of BoNT subtypes remaining as untapped potential sources for unique pharmaceuticals. Here, two recombinant hybrid toxins were engineered, consisting of domains from two BoNT subtypes that possess distinct duration of action and activity in human neurons and mice. We define the functional domains responsible for BoNT action and demonstrate creation of functional hybrid BoNTs with new therapeutic potential, while remaining sensitive to antitoxins and therapies to native BoNT.Sabine PellettMarite BradshawWilliam H. TeppChristina L. PierRegina C. M. WhitemarshChen ChenJoseph T. BarbieriEric A. JohnsonAmerican Society for MicrobiologyarticleBoNTbotulinum neurotoxinduration of actionhybrid toxinsubtypeMicrobiologyQR1-502ENmBio, Vol 9, Iss 2 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	BoNT botulinum neurotoxin duration of action hybrid toxin subtype Microbiology QR1-502
spellingShingle	BoNT botulinum neurotoxin duration of action hybrid toxin subtype Microbiology QR1-502 Sabine Pellett Marite Bradshaw William H. Tepp Christina L. Pier Regina C. M. Whitemarsh Chen Chen Joseph T. Barbieri Eric A. Johnson The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes
description	ABSTRACT Botulinum neurotoxin (BoNT) is the causative agent of botulism and a widely used pharmaceutical to treat a variety of neurological diseases. BoNTs are 150-kDa protein toxins organized into heavy chain (HC) and light chain (LC) domains linked by a disulfide bond. The HC selectively binds to neurons and aids cell entry of the enzymatically active LC. There are seven immunological BoNT serotypes (A to G); each serotype includes genetic variants, termed subtypes. Only two subtypes, BoNT/A1 and BoNT/B1, are currently used as therapeutics. BoNT serotype A (BoNT/A) subtypes A2 to A8 show distinct potency, duration of action, and pathology relative to BoNT/A1. Specifically, BoNT/A3 possesses shorter duration of action and elicits distinct symptoms in mice at high toxin doses. In this report, we analyzed the roles of LC and HC of BoNT/A3 for duration of action, neuronal cell entry, and mouse pathology by using clostridium-derived recombinant hybrid BoNTs consisting of reciprocal LC and HC (BoNTA1/A3 and BoNTA3/A1). Hybrid toxins were processed in their expression host to a dichain BoNT consisting of LC and HC linked via a disulfide bond. The LC and HC defined BoNT potency in mice and BoNT toxicity for cultured neuronal cells, while the LC defined the duration of BoNT action in cell and mouse models. Protein alignment identified a previously unrecognized region within the LC subtype A3 (LC/A3) relative to the other LC serotype A (LC/A) subtypes (low primary acid homology [LPH]) that correlated to intracellular LC localization. This study shows the utility of recombinant hybrid BoNTs with new therapeutic potential, while remaining sensitive to antitoxins and therapies to native BoNT. IMPORTANCE Botulinum neurotoxins are the most potent protein toxins for humans and potential bioterrorism threats, but they are also widely used as pharmaceuticals. Within the large family of BoNTs, only two subtypes are currently used as pharmaceuticals, with a large number of BoNT subtypes remaining as untapped potential sources for unique pharmaceuticals. Here, two recombinant hybrid toxins were engineered, consisting of domains from two BoNT subtypes that possess distinct duration of action and activity in human neurons and mice. We define the functional domains responsible for BoNT action and demonstrate creation of functional hybrid BoNTs with new therapeutic potential, while remaining sensitive to antitoxins and therapies to native BoNT.
format	article
author	Sabine Pellett Marite Bradshaw William H. Tepp Christina L. Pier Regina C. M. Whitemarsh Chen Chen Joseph T. Barbieri Eric A. Johnson
author_facet	Sabine Pellett Marite Bradshaw William H. Tepp Christina L. Pier Regina C. M. Whitemarsh Chen Chen Joseph T. Barbieri Eric A. Johnson
author_sort	Sabine Pellett
title	The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes
title_short	The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes
title_full	The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes
title_fullStr	The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes
title_full_unstemmed	The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes
title_sort	light chain defines the duration of action of botulinum toxin serotype a subtypes
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/aedc52e97f8748b7906c9ad8d9dcb24e
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The Light Chain Defines the Duration of Action of Botulinum Toxin Serotype A Subtypes

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