BspA and Pmp proteins ofTrichomonas vaginalismediate adherence to host cells

AbstractTrichomonas vaginalisis one of the most widespread, sexually transmitted pathogens. The infection involves a morphological switch from a free-swimming pyriform trophozoite to an amoeboid cell upon adhesion to host epithelial cells. While details on how the switch is induced and to what proteins of the host surface the parasite adheres remain poorly characterized, several surface proteins of the parasite itself have been identified as potential candidates. Among those are two expanded protein families that harbor domains that share similarity to functionally investigated surface proteins of prokaryotic oral pathogens; these are the BspA proteins of Bacteroidales and Spirochaetales, and the Pmp proteins of Chlamydiales. We sequenced the transcriptomes of five Trichomonads and screened for the presence of BspA and Pmp domain-containing proteins and tested the ability of individualT. vaginaliscandidates to mediate adhesion. Here we demonstrate that (i) BspA and Pmp domain-containing proteins are specifically expanded inT. vaginalisin comparison to other Trichomonads, and that (ii) individual proteins of both families have the ability to increase adhesion performance in a non-virulentT. vaginalisstrain andTetratrichomonas gallinarum, a parasite usually known to infect birds but not humans. Our results initiate the functional characterization of these two broadly distributed protein families, whose origin we trace back to the origin of Trichomonads themselves.

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A Cell Surface Aggregation-Promoting Factor fromLactobacillus gasseriContributes to Inhibition ofTrichomonas vaginalisAdhesion to Human Vaginal Ectocervical Cells

Infection and Immunity ◽

10.1128/iai.00907-17 ◽

2018 ◽

Vol 86 (8) ◽

Cited By ~ 13

Author(s):

Niha Phukan ◽

Anna E. S. Brooks ◽

Augusto Simoes-Barbosa

Keyword(s):

Trichomonas Vaginalis ◽

Protozoan Parasite ◽

Surface Proteins ◽

Host Cells ◽

Protective Mechanisms ◽

Transmitted Infection ◽

Content Type ◽

Sexually Transmitted ◽

Vaginal Lactobacilli ◽

A Cell

ABSTRACTTrichomoniasis, a prevalent sexually transmitted infection, is commonly symptomatic in women. The causative agent isTrichomonas vaginalis, an extracellular protozoan parasite. The host-protective mechanisms and molecules of vaginal lactobacilli that counteract this pathogen are largely unknown. This study examines the inhibition promoted byLactobacillus gasseriagainst the adhesion ofT. vaginalisto host cells, a critical virulence aspect of this pathogen. We observed that the vaginal strainL. gasseriATCC 9857 is highly inhibitory by various contact-dependent mechanisms and that surface proteins are largely responsible for this inhibitory phenotype. We found that the aggregation-promoting factor APF-2 from these bacteria significantly contributes to inhibition of the adhesion ofT. vaginalisto human vaginal ectocervical cells. Understanding the molecules and mechanisms used by lactobacilli to protect the host againstT. vaginalismight help in the development of novel and specific therapeutic strategies that take advantage of the natural microbiota.

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Functional Characterization of a Redundant Plasmodium TRAP Family Invasin, TRAP-Like Protein, by Aldolase Binding and a Genetic Complementation Test

Eukaryotic Cell ◽

10.1128/ec.00089-08 ◽

2008 ◽

Vol 7 (6) ◽

pp. 1062-1070 ◽

Cited By ~ 39

Author(s):

Kirsten Heiss ◽

Hui Nie ◽

Sumit Kumar ◽

Thomas M. Daly ◽

Lawrence W. Bergman ◽

...

Keyword(s):

Functional Characterization ◽

Cell Entry ◽

Host Cells ◽

Complementation Test ◽

Type I ◽

Cycle Progression ◽

Targeted Gene Disruption ◽

Specific Host ◽

Host Cell Entry

ABSTRACT Efficient and specific host cell entry is of exquisite importance for intracellular pathogens. Parasites of the phylum Apicomplexa are highly motile and actively enter host cells. These functions are mediated by type I transmembrane invasins of the TRAP family that link an extracellular recognition event to the parasite actin-myosin motor machinery. We systematically tested potential parasite invasins for binding to the actin bridging molecule aldolase and complementation of the vital cytoplasmic domain of the sporozoite invasin TRAP. We show that the ookinete invasin CTRP and a novel, structurally related protein, termed TRAP-like protein (TLP), are functional members of the TRAP family. Although TLP is expressed in invasive stages, targeted gene disruption revealed a nonvital role during life cycle progression. This is the first genetic analysis of TLP, encoding a redundant TRAP family invasin, in the malaria parasite.

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Characteristics of a Series of Three Bacteriophages Infecting Salmonella enterica Strains

International Journal of Molecular Sciences ◽

10.3390/ijms21176152 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6152 ◽

Cited By ~ 1

Author(s):

Katarzyna Kosznik-Kwaśnicka ◽

Karolina Ciemińska ◽

Michał Grabski ◽

Łukasz Grabowski ◽

Marcin Górniak ◽

...

Keyword(s):

Salmonella Enterica ◽

Phage Therapy ◽

Functional Characterization ◽

Bacterial Biofilm ◽

Host Cells ◽

Detailed Characterization ◽

Food Protection ◽

Adsorption Rates ◽

Potential Use

Molecular and functional characterization of a series of three bacteriophages, vB_SenM-1, vB_SenM-2, and vB_SenS-3, infecting various Salmonella enterica serovars and strains is presented. All these phages were able to develop lytically while not forming prophages. Moreover, they were able to survive at pH 3. The phages revealed different host ranges within serovars and strains of S. enterica, different adsorption rates on host cells, and different lytic growth kinetics at various temperatures (in the range of 25 to 42 °C). They efficiently reduced the number of cells in the bacterial biofilm and decreased the biofilm mass. Whole genome sequences of these phages have been determined and analyzed, including their phylogenetic relationships. In conclusion, we have demonstrated detailed characterization of a series of three bacteriophages, vB_SenM-1, vB_SenM-2, and vB_SenS-3, which reveal favorable features in light of their potential use in phage therapy of humans and animals, as well as for food protection purposes.

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A Novel Trichomonas vaginalis Surface Protein Modulates Parasite Attachment via Protein:Host Cell Proteoglycan Interaction

mBio ◽

10.1128/mbio.03374-20 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Brenda M. Molgora ◽

Anand Kumar Rai ◽

Michael J. Sweredoski ◽

Annie Moradian ◽

Sonja Hess ◽

...

Keyword(s):

Host Cell ◽

Trichomonas Vaginalis ◽

Surface Protein ◽

Host Cells ◽

Parasite Line ◽

Cell Adherence ◽

Common Component ◽

Content Type ◽

Sexually Transmitted ◽

Significant Difference

ABSTRACT Trichomonas vaginalis is a highly prevalent, sexually transmitted parasite which adheres to mucosal epithelial cells to colonize the human urogenital tract. Despite adherence being crucial for this extracellular parasite to thrive within the host, relatively little is known about the mechanisms or key molecules involved in this process. Here, we have identified and characterized a T. vaginalis hypothetical protein, TVAG_157210 (TvAD1), as a surface protein that plays an integral role in parasite adherence to the host. Quantitative proteomics revealed TvAD1 to be ∼4-fold more abundant in parasites selected for increased adherence (MA parasites) than the isogenic parental (P) parasite line. De novo modeling suggested that TvAD1 binds N-acetylglucosamine (GlcNAc), a sugar comprising host glycosaminoglycans (GAGs). Adherence assays utilizing GAG-deficient cell lines determined that host GAGs, primarily heparan sulfate (HS), mediate adherence of MA parasites to host cells. TvAD1 knockout (KO) parasites, generated using CRISPR-Cas9, were found to be significantly reduced in host cell adherence, a phenotype that is rescued by overexpression of TvAD1 in KO parasites. In contrast, there was no significant difference in parasite adherence to GAG-deficient lines by KO parasites compared with wild-type, which is contrary to that observed for KO parasites overexpressing TvAD1. Isothermal titration calorimetric (ITC) analysis showed that TvAD1 binds to HS, indicating that TvAD1 mediates host cell adherence via HS interaction. In addition to characterizing the role of TvAD1 in parasite adherence, these studies reveal a role for host GAG molecules in T. vaginalis adherence. IMPORTANCE The ability of the sexually transmitted parasite Trichomonas vaginalis to adhere to its human host is critical for establishing and maintaining an infection. Yet how parasites adhere to host cells is poorly understood. In this study, we employed a novel adherence selection method to identify proteins involved in parasite adherence to the host. This method led to the identification of a protein, with no previously known function, that is more abundant in parasites with increased capacity to bind host cells. Bioinformatic modeling and biochemical analyses revealed that this protein binds a common component on the host cell surface proteoglycans. Subsequent creation of parasites that lack this protein directly demonstrated that the protein mediates parasite adherence via an interaction with host cell proteoglycans. These findings both demonstrate a role for this protein in T. vaginalis adherence to the host and shed light on host cell molecules that participate in parasite colonization.

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Trichomonas vaginalis: current understanding of host–parasite interactions

Essays in Biochemistry ◽

10.1042/bse0510161 ◽

2011 ◽

Vol 51 ◽

pp. 161-175 ◽

Cited By ~ 58

Author(s):

Christopher M. Ryan ◽

Natalia de Miguel ◽

Patricia J. Johnson

Keyword(s):

Drug Targets ◽

Trichomonas Vaginalis ◽

Molecular Data ◽

Host Cells ◽

Current Status ◽

Rational Approach ◽

Sexually Transmitted ◽

Host Parasite Interactions ◽

Future Challenges ◽

Host Parasite

Trichomonas vaginalis is a sexually transmitted obligate extracellular parasite that colonizes the human urogenital tract. Despite being of critical importance to the parasite's survival relatively little is known about the mechanisms employed by T. vaginalis to establish an infection and thrive within its host. Several studies have focused on the interaction of the parasite with host cells and extracellular matrix, identifying multiple suspected T. vaginalis adhesins. However, with the exception of its surface lipophosphoglycan, the evidence supporting a role in adhesion is indirect or controversial for many candidate molecules. The availability of the T. vaginalis genome sequence paved the way for genomic analyses to search for proteins possibly involved in host–parasite interactions. Several proteomic analyses have also provided insight into surface, soluble and secreted proteins that may be involved in Trichomonas pathogenesis. Although the accumulation of molecular data allows for a more rational approach towards identifying drug targets and vaccine candidates for this medically important parasite, a continued effort is required to advance our understanding of its biology. In the present chapter, we review the current status of research aimed at understanding T. vaginalis pathogenesis. Applied experimental approaches, an overview of significant conclusions drawn from this research and future challenges are discussed.

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Quantitative proteomic analysis and functional characterization of Acanthamoeba castellanii exosome-like vesicles

Parasites & Vectors ◽

10.1186/s13071-019-3725-z ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 4

Author(s):

Wei-Chen Lin ◽

Chia-Yun Tsai ◽

Jian-Ming Huang ◽

Shang-Rung Wu ◽

Lichieh Julie Chu ◽

...

Keyword(s):

Immune Response ◽

Proteomic Analysis ◽

Functional Characterization ◽

Acanthamoeba Castellanii ◽

Host Cells ◽

C6 Cells ◽

Proteomic Profiling ◽

Quantitative Proteomic Analysis ◽

Rat Glioma

Abstract Background Pathogenic protozoans use extracellular vesicles (EVs) for intercellular communication and host manipulation. Acanthamoeba castellanii is a free-living protozoan that may cause severe keratitis and fatal granulomatous encephalitis. Although several secreted molecules have been shown to play crucial roles in the pathogenesis of Acanthamoeba, the functions and components of parasite-derived EVs are far from understood. Methods Purified EVs from A. castellanii were confirmed by electron microscopy and nanoparticle tracking analysis. The functional roles of parasite-derived EVs in the cytotoxicity to and immune response of host cells were examined. The protein composition in EVs from A. castellanii was identified and quantified by LC-MS/MS analysis. Results EVs from A. castellanii fused with rat glioma C6 cells. The parasite-derived EVs induced an immune response from human THP-1 cells and a cytotoxic effect in C6 cells. Quantitative proteomic analysis identified a total of 130 proteins in EVs. Among the identified proteins, hydrolases (50.2%) and oxidoreductases (31.7%) were the largest protein families in EVs. Furthermore, aminopeptidase activities were confirmed in EVs from A. castellanii. Conclusions The proteomic profiling and functional characterization of EVs from A. castellanii provide an in-depth understanding of the molecules packaged into EVs and their potential mechanisms mediating the pathogenesis of this parasite.

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Functional Characterization of Borrelia spielmanii Outer Surface Proteins That Interact with Distinct Members of the Human Factor H Protein Family and with Plasminogen

Infection and Immunity ◽

10.1128/iai.00691-09 ◽

2009 ◽

Vol 78 (1) ◽

pp. 39-48 ◽

Cited By ~ 24

Author(s):

Annekatrin Seling ◽

Corinna Siegel ◽

Volker Fingerle ◽

Brandon L. Jutras ◽

Catherine A. Brissette ◽

...

Keyword(s):

Outer Surface ◽

Functional Characterization ◽

Protein Family ◽

Factor H ◽

Surface Proteins ◽

Single Amino Acid ◽

Related Protein ◽

Outer Surface Proteins ◽

Complement Regulator

ABSTRACT Acquisition of complement regulator factor H (CFH) and factor H-like protein 1 (CFHL1) from human serum enables Borrelia spielmanii, one of the etiological agents of Lyme disease, to evade complement-mediated killing by the human host. Up to three distinct complement regulator-acquiring surface proteins (CRASPs) may be expressed by serum-resistant B. spielmanii, each exhibiting an affinity for CFH and/or CFHL1. Here, we describe the functional characterization of the 15-kDa CRASPs of B. spielmanii, members of the polymorphic Erp (OspE/F-related) protein family, that bind two distinct host complement regulators, CFH and factor H-related protein 1 (CFHR1), but not CFHL1. CFH bound to the B. spielmanii CRASPs maintained cofactor activity for factor I-mediated C3b inactivation. Three naturally occurring alleles of this protein bound CFH and CFHR1 while a fourth natural allele could not. Comparative sequence analysis of these protein alleles identified a single amino acid, histidine-79, as playing a significant role in CFH/CFHR1 binding, with substitution by an arginine completely abrogating ligand binding. The mutation of His-79 to Arg did not inhibit binding of plasminogen, another known ligand of this group of borrelial outer-surface proteins.

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