Changing the Receptor Specificity of Anthrax Toxin

ABSTRACT The actions of many bacterial toxins depend on their ability to bind to one or more cell-surface receptors. Anthrax toxin acts by a sequence of events that begins when the protective-antigen (PA) moiety of the toxin binds to either one of two cell-surface proteins, ANTXR1 and ANTXR2, and is...

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Autores principales:	Adva Mechaly, Andrew J. McCluskey, R. John Collier
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/41d69d926d1248ddba00e28a2290cfc0
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spelling	oai:doaj.org-article:41d69d926d1248ddba00e28a2290cfc02021-11-15T15:39:02ZChanging the Receptor Specificity of Anthrax Toxin10.1128/mBio.00088-122150-7511https://doaj.org/article/41d69d926d1248ddba00e28a2290cfc02012-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00088-12https://doaj.org/toc/2150-7511ABSTRACT The actions of many bacterial toxins depend on their ability to bind to one or more cell-surface receptors. Anthrax toxin acts by a sequence of events that begins when the protective-antigen (PA) moiety of the toxin binds to either one of two cell-surface proteins, ANTXR1 and ANTXR2, and is proteolytically activated. The activated PA self-associates to form oligomeric pore precursors, which, in turn, bind the enzymatic moieties of the toxin and transport them to the cytosol. We introduced a double mutation into domain 4 of PA to ablate its native receptor-binding function and fused epidermal growth factor (EGF) to the C terminus of the mutated protein. The resulting fusion protein transported enzymatic effector proteins into a cell line that expressed the EGF receptor (A431 cells), but not into a line lacking this receptor (CHO-K1 cells). Addition of excess free EGF blocked transport of effector proteins into A431 cells via the fusion protein, but not via native PA. We also showed that fusing the diphtheria toxin receptor-binding domain to the C terminus of the mutated PA channeled effector-protein transport through the diphtheria toxin receptor. PA fusion proteins with altered receptor specificity may be useful in biological research and could have practical applications, including ablation or perturbation of selected populations of cells in vivo. IMPORTANCE Bacterial toxins that act within mammalian cells have receptor-dependent mechanisms to transport their enzymatic components to the cytoplasmic compartment. By inactivating or otherwise modifying their respective intracellular targets, these intracellular effectors disrupt metabolic pathways and in some cases cause death of the cell. Our results show that the receptor specificity of the transport protein of anthrax toxin may be readily changed, raising the possibility that receptor-redirected forms of protective antigen (PA) and PA homologs may be useful for research and medical applications requiring modification or ablation of designated populations of cells.Adva MechalyAndrew J. McCluskeyR. John CollierAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 3 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Adva Mechaly Andrew J. McCluskey R. John Collier Changing the Receptor Specificity of Anthrax Toxin
description	ABSTRACT The actions of many bacterial toxins depend on their ability to bind to one or more cell-surface receptors. Anthrax toxin acts by a sequence of events that begins when the protective-antigen (PA) moiety of the toxin binds to either one of two cell-surface proteins, ANTXR1 and ANTXR2, and is proteolytically activated. The activated PA self-associates to form oligomeric pore precursors, which, in turn, bind the enzymatic moieties of the toxin and transport them to the cytosol. We introduced a double mutation into domain 4 of PA to ablate its native receptor-binding function and fused epidermal growth factor (EGF) to the C terminus of the mutated protein. The resulting fusion protein transported enzymatic effector proteins into a cell line that expressed the EGF receptor (A431 cells), but not into a line lacking this receptor (CHO-K1 cells). Addition of excess free EGF blocked transport of effector proteins into A431 cells via the fusion protein, but not via native PA. We also showed that fusing the diphtheria toxin receptor-binding domain to the C terminus of the mutated PA channeled effector-protein transport through the diphtheria toxin receptor. PA fusion proteins with altered receptor specificity may be useful in biological research and could have practical applications, including ablation or perturbation of selected populations of cells in vivo. IMPORTANCE Bacterial toxins that act within mammalian cells have receptor-dependent mechanisms to transport their enzymatic components to the cytoplasmic compartment. By inactivating or otherwise modifying their respective intracellular targets, these intracellular effectors disrupt metabolic pathways and in some cases cause death of the cell. Our results show that the receptor specificity of the transport protein of anthrax toxin may be readily changed, raising the possibility that receptor-redirected forms of protective antigen (PA) and PA homologs may be useful for research and medical applications requiring modification or ablation of designated populations of cells.
format	article
author	Adva Mechaly Andrew J. McCluskey R. John Collier
author_facet	Adva Mechaly Andrew J. McCluskey R. John Collier
author_sort	Adva Mechaly
title	Changing the Receptor Specificity of Anthrax Toxin
title_short	Changing the Receptor Specificity of Anthrax Toxin
title_full	Changing the Receptor Specificity of Anthrax Toxin
title_fullStr	Changing the Receptor Specificity of Anthrax Toxin
title_full_unstemmed	Changing the Receptor Specificity of Anthrax Toxin
title_sort	changing the receptor specificity of anthrax toxin
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/41d69d926d1248ddba00e28a2290cfc0
work_keys_str_mv	AT advamechaly changingthereceptorspecificityofanthraxtoxin AT andrewjmccluskey changingthereceptorspecificityofanthraxtoxin AT rjohncollier changingthereceptorspecificityofanthraxtoxin
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Changing the Receptor Specificity of Anthrax Toxin

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