Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain

Huang, Jian; Lin, Wei Chen; Li, Sung Chou; Shih, Ming Chang; Chan, Wen; Shin, Jyh Wei; Huang, Fu Chin

doi:10.1016/j.exppara.2016.03.018

Cited by 11 publications

(11 citation statements)

References 55 publications

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“…In this study, the results showed that Asp more rapidly hydrolyzed Leu-AMC than Arg-AMC, as well as the recombinant M28AP ( Figure 2B). These results aligned with the secreted proteomics analysis, which proved that Asp contain the M28AP, because the recombinant M28AP hydrolyzed Leu-AMC, which matches the Asp hydrolyzed Leu-AMC subtracts [25]. The normal pH range of human tears is 6.5 to 7.6; the mean value is 7.0 [32].…”

Section: Biochemical Characterization Of M28apsupporting

confidence: 75%

“…In a previous study, we established the extracellular secreted proteomic database of Acanthamoeba spp. [25]. We found a novel protein secreted by Acanthamoeba spp., M28AP, from the extracellular secreted proteomic database as a target of the human innate immune defense system [26].…”

Section: Biochemical Characterization Of M28apmentioning

confidence: 88%

“…We previously showed that the M20/M25/M40 superfamily aminopeptidase protein, which is an aminopeptidase found in Acanthamoeba-secreted proteins (Asp), causes cell damage and disruption [24]. We also identified a novel protein that was secreted by Acanthamoeba spp., an M28 aminopeptidase (M28AP), as a target of the human innate immune defense [25,26]. However, little is known regarding the biochemical and functional characterization of M28AP.…”

Section: Introductionmentioning

confidence: 99%

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The Biochemical and Functional Characterization of M28 Aminopeptidase Protein Secreted by Acanthamoeba spp. on Host Cell Interaction

2019

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Acanthamoeba are a free-living protozoan whose pathogenic strain can cause severe human diseases, such as granulomatous encephalitis and keratitis. As such, the pathogenic mechanism between humans and Acanthamoeba is still unknown. In our previous study, we identified the secreted Acanthamoeba M28 aminopeptidase (M28AP) and then suggested that M28AP can degrade human C3b and iC3b for inhibiting the destruction of Acanthamoeba spp. with the human immune response. We constructed the produced the recombinant M28AP from a CHO cell, which is a mammalian expression system, to characterize the biochemical properties of Acanthamoeba M28AP. The recombinant M28AP more rapidly hydrolyzed Leu-AMC than Arg-AMC and could be inhibited by EDTA treatment. We show that recombinant M28AP can be delivered into the individual cell line and cause cell line apoptosis in a co-culture model. In conclusion, we successfully investigated the potential molecular characteristics of M28AP.

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Section: Biochemical Characterization Of M28apsupporting

confidence: 75%

Section: Biochemical Characterization Of M28apmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 99%

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The Biochemical and Functional Characterization of M28 Aminopeptidase Protein Secreted by Acanthamoeba spp. on Host Cell Interaction

2019

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“…Results show that there was no significant change in C6 cell coverage area between C6 cells which were co-incubated with the non-pathogenic ATCC_30010 strain and control (mock) ( Figure 1 ). The live pathogenic Acanthamoeba protists, NCKH_D and ATCC_50492, could cause C6 cell damage after co-incubation for 24 h. In our previous study, we have established an RNA-seq database and a comparative proteomic analysis of extracellular secreted proteins, both of them including the ATCC 30010 and the NCKH_D strains [ 16 ]. In order to gain a deeper understanding of the pathogenesis mechanism, we focus on the cytopathic effect process of NCKH_D.…”

Section: Resultsmentioning

confidence: 99%

“…In our previous study, we reported that datasets of secreted proteomic Acanthamoeba strains, including the non-pathogenic ATCC_30010 strain and clinically pathogenic isolates, had been established established [ 16 ]. These clinically pathogenic Acanthamoeba isolates were collected by National Cheng Kung University (NCKU) Hospital from patients infected with AK [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

Pathogenic Acanthamoeba castellanii Secretes the Extracellular Aminopeptidase M20/M25/M40 Family Protein to Target Cells for Phagocytosis by Disruption

et al. 2017

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Acanthamoeba is free-living protist pathogen capable of causing a blinding keratitis and granulomatous encephalitis. However, the mechanisms of Acanthamoeba pathogenesis are still not clear. Here, our results show that cells co-cultured with pathogenic Acanthamoeba would be spherical and floated, even without contacting the protists. Then, the Acanthamoeba protists would contact and engulf these cells. In order to clarify the contact-independent pathogenesis mechanism in Acanthamoeba, we collected the Acanthamoeba-secreted proteins (Asp) to incubate with cells for identifying the extracellular virulent factors and investigating the cytotoxicity process. The Asps of pathogenic Acanthamoeba express protease activity to reactive Leu amino acid in ECM and induce cell-losing adhesion ability. The M20/M25/M40 superfamily aminopeptidase protein (ACA1_264610), an aminopeptidase be found in Asp, is upregulated after Acanthamoeba and C6 cell co-culturing for 6 h. Pre-treating the Asp with leucine aminopeptidase inhibitor and the specific antibodies of Acanthamoeba M20/M25/M40 superfamily aminopeptidase could reduce the cell damage during Asp and cell co-incubation. These results suggest an important functional role of the Acanthamoeba secreted extracellular aminopeptidases in the Acanthamoeba pathogenesis process. This study provides information regarding clinically pathogenic isolates to target specific molecules and design combined drugs.

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Protein profiling of Acanthamoeba species using MALDI-TOF MS for specific identification of Acanthamoeba genotype

et al. 2018

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Acanthamoeba spp. are ubiquitous in the environment and have the potential to cause severe infections. The different genotypes of Acanthamoeba have been shown to influence the severity of the disease and response to therapy. Characterizing Acanthamoeba spp. upto genotype can aid in infection control practices. Twenty-five Acanthamoeba isolates, characterized by 18S rDNA sequencing, were subjected to MALDI-TOF MS analysis by creating a database for the individual genotypes. The differentiating features of the various spectra were observed; the coded samples were then tested against the created database. The results of identification were compared with sequencing. Five different genotypes were obtained-T3, T4, T5, T10, and T11. Spectral analysis revealed genus-specific and genotype-specific peaks. The peak patterns for individual genotype were discrete and reproducible. Clinical isolates produced different peaks from the environmental isolate of the same genotype. A concordance of 92% was obtained with MALDI-TOF MS in comparison with 18sDNA sequencing. MALDI-TOF MS, once optimized, has the potential to reliably identify the genotype of Acanthamoeba spp. and to differentiate clinical isolate from mere contaminant.

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Comparative proteomic analysis of extracellular secreted proteins expressed by two pathogenic Acanthamoeba castellanii clinical isolates and a non-pathogenic ATCC strain

Cited by 11 publications

References 55 publications

The Biochemical and Functional Characterization of M28 Aminopeptidase Protein Secreted by Acanthamoeba spp. on Host Cell Interaction

The Biochemical and Functional Characterization of M28 Aminopeptidase Protein Secreted by Acanthamoeba spp. on Host Cell Interaction

Pathogenic Acanthamoeba castellanii Secretes the Extracellular Aminopeptidase M20/M25/M40 Family Protein to Target Cells for Phagocytosis by Disruption

Protein profiling of Acanthamoeba species using MALDI-TOF MS for specific identification of Acanthamoeba genotype

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