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Functionally Expression of Metalloproteinase in Taenia solium Metacestode and Its Evaluation for Serodiagnosis of Cysticercosis.

Zhang Y, Bae YA, Zong HY, Kong Y, Cai GB - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed.The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates.TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Genetics, Wuhan University School of Basic Medicial Sciences, Wuhan 430071, China.

ABSTRACT

Background: Parasite proteases have important roles in cleavage of host proteins during the invasion of host tissues and participate in the parasite's evasion from the host's immune response. The aim of the present study was to estimate a metalloproteinase properties of Taenia solium metacestode (TsMP) during host-parasite interactions, and evaluate its potential as a serodiagnostic antigen for cysticercosis.

Methods: The cDNA coding for the mature catalytic domain of TsMP was cloned into pGEX-6P-1 expression vector. A recombinant glutathione S-transferase and TsMP fusion protein was induced. After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed. Immunoblot assay was processed to evaluate its potential as a serodiagnostic antigen for cysticercosis.

Results: The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates. In immunoblot assay, 87.5% of sera from patients with cysticercosis showed strong reactivity. In sera from patients with other parasitic infections and from normal controls, it showed high specificity.

Conclusions: TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle. A single recombinant TsMP antigen could have a potential value for serodiagnosis of cysticercosis.

No MeSH data available.


Related in: MedlinePlus

Degradation of various host proteins by the purified recombinant mTsMP enzyme. Each macromolecular protein substrate (10 µg) including Collagen, Fibronectin, Bovine serum albumin (BSA) and Haemoglobin was incubated with recombinant mTsMP (1 µg) in 50 mM Tris-HCl buffer (pH 7.5) for 2 h or 4 h at 37°C and analyzed by 10% SDS-PAGE. Lane C, control (protein without recombinant mTsMP); lane1 and lane 2, incubated with recombinant mTsMP for 2 and 4 h respectively
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Figure 4: Degradation of various host proteins by the purified recombinant mTsMP enzyme. Each macromolecular protein substrate (10 µg) including Collagen, Fibronectin, Bovine serum albumin (BSA) and Haemoglobin was incubated with recombinant mTsMP (1 µg) in 50 mM Tris-HCl buffer (pH 7.5) for 2 h or 4 h at 37°C and analyzed by 10% SDS-PAGE. Lane C, control (protein without recombinant mTsMP); lane1 and lane 2, incubated with recombinant mTsMP for 2 and 4 h respectively

Mentions: To investigate the putative biological roles of TsMP, the proteolytic activity of recombinant mTsMP against several host proteins including collagen, fibronectin, BSA and haemoglobin was determined. All protein substrates except haemoglobin used in this study were hydrolysis by the recombinant mTsMP proteinase at neutral pH. However, the degradation of haemoglobin was not degraded significantly (Fig. 4).


Functionally Expression of Metalloproteinase in Taenia solium Metacestode and Its Evaluation for Serodiagnosis of Cysticercosis.

Zhang Y, Bae YA, Zong HY, Kong Y, Cai GB - Iran J Parasitol (2016 Jan-Mar)

Degradation of various host proteins by the purified recombinant mTsMP enzyme. Each macromolecular protein substrate (10 µg) including Collagen, Fibronectin, Bovine serum albumin (BSA) and Haemoglobin was incubated with recombinant mTsMP (1 µg) in 50 mM Tris-HCl buffer (pH 7.5) for 2 h or 4 h at 37°C and analyzed by 10% SDS-PAGE. Lane C, control (protein without recombinant mTsMP); lane1 and lane 2, incubated with recombinant mTsMP for 2 and 4 h respectively
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835468&req=5

Figure 4: Degradation of various host proteins by the purified recombinant mTsMP enzyme. Each macromolecular protein substrate (10 µg) including Collagen, Fibronectin, Bovine serum albumin (BSA) and Haemoglobin was incubated with recombinant mTsMP (1 µg) in 50 mM Tris-HCl buffer (pH 7.5) for 2 h or 4 h at 37°C and analyzed by 10% SDS-PAGE. Lane C, control (protein without recombinant mTsMP); lane1 and lane 2, incubated with recombinant mTsMP for 2 and 4 h respectively
Mentions: To investigate the putative biological roles of TsMP, the proteolytic activity of recombinant mTsMP against several host proteins including collagen, fibronectin, BSA and haemoglobin was determined. All protein substrates except haemoglobin used in this study were hydrolysis by the recombinant mTsMP proteinase at neutral pH. However, the degradation of haemoglobin was not degraded significantly (Fig. 4).

Bottom Line: After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed.The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates.TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Genetics, Wuhan University School of Basic Medicial Sciences, Wuhan 430071, China.

ABSTRACT

Background: Parasite proteases have important roles in cleavage of host proteins during the invasion of host tissues and participate in the parasite's evasion from the host's immune response. The aim of the present study was to estimate a metalloproteinase properties of Taenia solium metacestode (TsMP) during host-parasite interactions, and evaluate its potential as a serodiagnostic antigen for cysticercosis.

Methods: The cDNA coding for the mature catalytic domain of TsMP was cloned into pGEX-6P-1 expression vector. A recombinant glutathione S-transferase and TsMP fusion protein was induced. After refolding and purification, enzymatic properties of the recombinant metalloproteinase were observed. Immunoblot assay was processed to evaluate its potential as a serodiagnostic antigen for cysticercosis.

Results: The recombinant TsMP protein showed proteolytic activity, which preferred host extracellular matrix proteins such as collagen and fibronectin as degradable substrates. In immunoblot assay, 87.5% of sera from patients with cysticercosis showed strong reactivity. In sera from patients with other parasitic infections and from normal controls, it showed high specificity.

Conclusions: TsMP might be involved in the processing of numerous host proteins and play an important role in the parasite life cycle. A single recombinant TsMP antigen could have a potential value for serodiagnosis of cysticercosis.

No MeSH data available.


Related in: MedlinePlus