Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection

ABSTRACT Diverse intracellular pathogens rely on eukaryotic cell surface disulfide reductases to invade host cells. Pharmacologic inhibition of these enzymes is cytotoxic, making it impractical for treatment. Identifying and mechanistically dissecting microbial proteins that co-opt surface reductase...

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Autores principales:	Ryan S. Green, Waheeda A. Naimi, Lee D. Oliver, Nathaniel O’Bier, Jaehyung Cho, Daniel H. Conrad, Rebecca K. Martin, Richard T. Marconi, Jason A. Carlyon
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	adhesin Anaplasma phagocytophilum obligate intracellular bacteria protein disulfide isomerase host-pathogen interactions Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5fcfb0454aa64878b57f1f16d651b6f8
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spelling	oai:doaj.org-article:5fcfb0454aa64878b57f1f16d651b6f82021-11-15T15:56:58ZBinding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection10.1128/mBio.03141-192150-7511https://doaj.org/article/5fcfb0454aa64878b57f1f16d651b6f82020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.03141-19https://doaj.org/toc/2150-7511ABSTRACT Diverse intracellular pathogens rely on eukaryotic cell surface disulfide reductases to invade host cells. Pharmacologic inhibition of these enzymes is cytotoxic, making it impractical for treatment. Identifying and mechanistically dissecting microbial proteins that co-opt surface reductases could reveal novel targets for disrupting this common infection strategy. Anaplasma phagocytophilum invades neutrophils by an incompletely defined mechanism to cause the potentially fatal disease granulocytic anaplasmosis. The bacterium’s adhesin, Asp14, contributes to invasion by virtue of its C terminus engaging an unknown receptor. Yeast-two hybrid analysis identified protein disulfide isomerase (PDI) as an Asp14 binding partner. Coimmunoprecipitation confirmed the interaction and validated it to be Asp14 C terminus dependent. PDI knockdown and antibody-mediated inhibition of PDI reductase activity impaired A. phagocytophilum infection of but not binding to host cells. Infection during PDI inhibition was rescued when the bacterial but not host cell surface disulfide bonds were chemically reduced with tris(2-carboxyethyl)phosphine-HCl (TCEP). TCEP also restored bacterial infectivity in the presence of an Asp14 C terminus blocking antibody that otherwise inhibits infection. A. phagocytophilum failed to productively infect myeloid-specific-PDI conditional-knockout mice, marking the first demonstration of in vivo microbial dependency on PDI for infection. Mutational analyses identified the Asp14 C-terminal residues that are critical for binding PDI. Thus, Asp14 binds and brings PDI proximal to A. phagocytophilum surface disulfide bonds that it reduces, which enables cellular and in vivo infection. IMPORTANCE Anaplasma phagocytophilum infects neutrophils to cause granulocytic anaplasmosis, an emerging potentially fatal disease and the second-most common tick-borne illness in the United States. Treatment options are limited, and no vaccine exists. Due to the bacterium’s obligatory intracellular lifestyle, A. phagocytophilum survival and pathogenesis are predicated on its ability to enter host cells. Understanding its invasion mechanism will yield new targets for preventing bacterial entry and, hence, disease. We report a novel entry pathway in which the A. phagocytophilum outer membrane protein Asp14 binds host cell surface protein disulfide isomerase via specific C-terminal residues to promote reduction of bacterial surface disulfide bonds, which is critical for cellular invasion and productive infection in vivo. Targeting the Asp14 C terminus could be used to prevent/treat granulocytic anaplasmosis. Our findings have broad implications, as a thematically similar approach could be applied to block infection by other intracellular microbes that exploit cell surface reductases.Ryan S. GreenWaheeda A. NaimiLee D. OliverNathaniel O’BierJaehyung ChoDaniel H. ConradRebecca K. MartinRichard T. MarconiJason A. CarlyonAmerican Society for MicrobiologyarticleadhesinAnaplasma phagocytophilumobligate intracellular bacteriaprotein disulfide isomerasehost-pathogen interactionsMicrobiologyQR1-502ENmBio, Vol 11, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	adhesin Anaplasma phagocytophilum obligate intracellular bacteria protein disulfide isomerase host-pathogen interactions Microbiology QR1-502
spellingShingle	adhesin Anaplasma phagocytophilum obligate intracellular bacteria protein disulfide isomerase host-pathogen interactions Microbiology QR1-502 Ryan S. Green Waheeda A. Naimi Lee D. Oliver Nathaniel O’Bier Jaehyung Cho Daniel H. Conrad Rebecca K. Martin Richard T. Marconi Jason A. Carlyon Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection
description	ABSTRACT Diverse intracellular pathogens rely on eukaryotic cell surface disulfide reductases to invade host cells. Pharmacologic inhibition of these enzymes is cytotoxic, making it impractical for treatment. Identifying and mechanistically dissecting microbial proteins that co-opt surface reductases could reveal novel targets for disrupting this common infection strategy. Anaplasma phagocytophilum invades neutrophils by an incompletely defined mechanism to cause the potentially fatal disease granulocytic anaplasmosis. The bacterium’s adhesin, Asp14, contributes to invasion by virtue of its C terminus engaging an unknown receptor. Yeast-two hybrid analysis identified protein disulfide isomerase (PDI) as an Asp14 binding partner. Coimmunoprecipitation confirmed the interaction and validated it to be Asp14 C terminus dependent. PDI knockdown and antibody-mediated inhibition of PDI reductase activity impaired A. phagocytophilum infection of but not binding to host cells. Infection during PDI inhibition was rescued when the bacterial but not host cell surface disulfide bonds were chemically reduced with tris(2-carboxyethyl)phosphine-HCl (TCEP). TCEP also restored bacterial infectivity in the presence of an Asp14 C terminus blocking antibody that otherwise inhibits infection. A. phagocytophilum failed to productively infect myeloid-specific-PDI conditional-knockout mice, marking the first demonstration of in vivo microbial dependency on PDI for infection. Mutational analyses identified the Asp14 C-terminal residues that are critical for binding PDI. Thus, Asp14 binds and brings PDI proximal to A. phagocytophilum surface disulfide bonds that it reduces, which enables cellular and in vivo infection. IMPORTANCE Anaplasma phagocytophilum infects neutrophils to cause granulocytic anaplasmosis, an emerging potentially fatal disease and the second-most common tick-borne illness in the United States. Treatment options are limited, and no vaccine exists. Due to the bacterium’s obligatory intracellular lifestyle, A. phagocytophilum survival and pathogenesis are predicated on its ability to enter host cells. Understanding its invasion mechanism will yield new targets for preventing bacterial entry and, hence, disease. We report a novel entry pathway in which the A. phagocytophilum outer membrane protein Asp14 binds host cell surface protein disulfide isomerase via specific C-terminal residues to promote reduction of bacterial surface disulfide bonds, which is critical for cellular invasion and productive infection in vivo. Targeting the Asp14 C terminus could be used to prevent/treat granulocytic anaplasmosis. Our findings have broad implications, as a thematically similar approach could be applied to block infection by other intracellular microbes that exploit cell surface reductases.
format	article
author	Ryan S. Green Waheeda A. Naimi Lee D. Oliver Nathaniel O’Bier Jaehyung Cho Daniel H. Conrad Rebecca K. Martin Richard T. Marconi Jason A. Carlyon
author_facet	Ryan S. Green Waheeda A. Naimi Lee D. Oliver Nathaniel O’Bier Jaehyung Cho Daniel H. Conrad Rebecca K. Martin Richard T. Marconi Jason A. Carlyon
author_sort	Ryan S. Green
title	Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection
title_short	Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection
title_full	Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection
title_fullStr	Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection
title_full_unstemmed	Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection
title_sort	binding of host cell surface protein disulfide isomerase by <named-content content-type="genus-species">anaplasma phagocytophilum</named-content> asp14 enables pathogen infection
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/5fcfb0454aa64878b57f1f16d651b6f8
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Binding of Host Cell Surface Protein Disulfide Isomerase by <named-content content-type="genus-species">Anaplasma phagocytophilum</named-content> Asp14 Enables Pathogen Infection

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