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Interaction of Leptospira elongation factor Tu with plasminogen and complement factor H: a metabolic leptospiral protein with moonlighting activities.

Wolff DG, Castiblanco-Valencia MM, Abe CM, Monaris D, Morais ZM, Souza GO, Vasconcellos SA, Isaac L, Abreu PA, Barbosa AS - PLoS ONE (2013)

Bottom Line: Known to perform more than one function at different times or in different places, it is found in several subcellular locations in a single organism, and may serve as a virulence factor in a range of important human pathogens.In this manner, EF-Tu may contribute to leptospiral tissue invasion and complement inactivation.To our knowledge, this is the first description of a leptospiral protein exhibiting moonlighting activities.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, Brasil.

ABSTRACT
The elongation factor Tu (EF-Tu), an abundant bacterial protein involved in protein synthesis, has been shown to display moonlighting activities. Known to perform more than one function at different times or in different places, it is found in several subcellular locations in a single organism, and may serve as a virulence factor in a range of important human pathogens. Here we demonstrate that Leptospira EF-Tu is surface-exposed and performs additional roles as a cell-surface receptor for host plasma proteins. It binds plasminogen in a dose-dependent manner, and lysine residues are critical for this interaction. Bound plasminogen is converted to active plasmin, which, in turn, is able to cleave the natural substrates C3b and fibrinogen. Leptospira EF-Tu also acquires the complement regulator Factor H (FH). FH bound to immobilized EF-Tu displays cofactor activity, mediating C3b degradation by Factor I (FI). In this manner, EF-Tu may contribute to leptospiral tissue invasion and complement inactivation. To our knowledge, this is the first description of a leptospiral protein exhibiting moonlighting activities.

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EF-Tu-bound plasminogen is converted to functionally active plasmin.Recombinant proteins or BSA (10 μg/mL), immobilized to microtiter plate wells, were incubated with plasminogen (20 μg/mL). After washing, uPA (3 U) and the chromogenic substrate D-valyl-leucyl-lysine-ρ-nitroanilide dihydrochloride (25µg/well) were added. Data represent the mean absorbance value at 405 nm ± the standard deviation of three independent experiments, each performed in duplicate. (* p < 0.05). .
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pone-0081818-g004: EF-Tu-bound plasminogen is converted to functionally active plasmin.Recombinant proteins or BSA (10 μg/mL), immobilized to microtiter plate wells, were incubated with plasminogen (20 μg/mL). After washing, uPA (3 U) and the chromogenic substrate D-valyl-leucyl-lysine-ρ-nitroanilide dihydrochloride (25µg/well) were added. Data represent the mean absorbance value at 405 nm ± the standard deviation of three independent experiments, each performed in duplicate. (* p < 0.05). .

Mentions: To assess if EF-Tu-bound plasminogen could be converted to active plasmin by exogenously supplied uPA, immobilized EF-Tu was incubated with plasminogen. After extensive washing, uPA and the chromogenic substrate D-valyl-leucyl-lysine-ρ-nitroanilide dihydrochloride were added. The newly generated plasmin was able to cleave the chromogenic substrate (Figure 4).


Interaction of Leptospira elongation factor Tu with plasminogen and complement factor H: a metabolic leptospiral protein with moonlighting activities.

Wolff DG, Castiblanco-Valencia MM, Abe CM, Monaris D, Morais ZM, Souza GO, Vasconcellos SA, Isaac L, Abreu PA, Barbosa AS - PLoS ONE (2013)

EF-Tu-bound plasminogen is converted to functionally active plasmin.Recombinant proteins or BSA (10 μg/mL), immobilized to microtiter plate wells, were incubated with plasminogen (20 μg/mL). After washing, uPA (3 U) and the chromogenic substrate D-valyl-leucyl-lysine-ρ-nitroanilide dihydrochloride (25µg/well) were added. Data represent the mean absorbance value at 405 nm ± the standard deviation of three independent experiments, each performed in duplicate. (* p < 0.05). .
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3842364&req=5

pone-0081818-g004: EF-Tu-bound plasminogen is converted to functionally active plasmin.Recombinant proteins or BSA (10 μg/mL), immobilized to microtiter plate wells, were incubated with plasminogen (20 μg/mL). After washing, uPA (3 U) and the chromogenic substrate D-valyl-leucyl-lysine-ρ-nitroanilide dihydrochloride (25µg/well) were added. Data represent the mean absorbance value at 405 nm ± the standard deviation of three independent experiments, each performed in duplicate. (* p < 0.05). .
Mentions: To assess if EF-Tu-bound plasminogen could be converted to active plasmin by exogenously supplied uPA, immobilized EF-Tu was incubated with plasminogen. After extensive washing, uPA and the chromogenic substrate D-valyl-leucyl-lysine-ρ-nitroanilide dihydrochloride were added. The newly generated plasmin was able to cleave the chromogenic substrate (Figure 4).

Bottom Line: Known to perform more than one function at different times or in different places, it is found in several subcellular locations in a single organism, and may serve as a virulence factor in a range of important human pathogens.In this manner, EF-Tu may contribute to leptospiral tissue invasion and complement inactivation.To our knowledge, this is the first description of a leptospiral protein exhibiting moonlighting activities.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Bacteriologia, Instituto Butantan, São Paulo, Brasil.

ABSTRACT
The elongation factor Tu (EF-Tu), an abundant bacterial protein involved in protein synthesis, has been shown to display moonlighting activities. Known to perform more than one function at different times or in different places, it is found in several subcellular locations in a single organism, and may serve as a virulence factor in a range of important human pathogens. Here we demonstrate that Leptospira EF-Tu is surface-exposed and performs additional roles as a cell-surface receptor for host plasma proteins. It binds plasminogen in a dose-dependent manner, and lysine residues are critical for this interaction. Bound plasminogen is converted to active plasmin, which, in turn, is able to cleave the natural substrates C3b and fibrinogen. Leptospira EF-Tu also acquires the complement regulator Factor H (FH). FH bound to immobilized EF-Tu displays cofactor activity, mediating C3b degradation by Factor I (FI). In this manner, EF-Tu may contribute to leptospiral tissue invasion and complement inactivation. To our knowledge, this is the first description of a leptospiral protein exhibiting moonlighting activities.

Show MeSH
Related in: MedlinePlus