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Leishmania mexicana: molecular cloning and characterization of enolase

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Leishmania mexicana : molecular cloning and characterization of enolase. / Quiñones, Wilfredo; Peña, Priscila; Domingo-Sananes, Maria; Cáceres, Ana; Michels, Paul A. M.; Avilan, Luisana; Concepción, Juan Luis.

In: Experimental Parasitology, Vol. 116, No. 3, 07.2007, p. 241-251.

Research output: Contribution to journalArticlepeer-review

Harvard

Quiñones, W, Peña, P, Domingo-Sananes, M, Cáceres, A, Michels, PAM, Avilan, L & Concepción, JL 2007, 'Leishmania mexicana: molecular cloning and characterization of enolase', Experimental Parasitology, vol. 116, no. 3, pp. 241-251. https://doi.org/10.1016/j.exppara.2007.01.008

APA

Quiñones, W., Peña, P., Domingo-Sananes, M., Cáceres, A., Michels, P. A. M., Avilan, L., & Concepción, J. L. (2007). Leishmania mexicana: molecular cloning and characterization of enolase. Experimental Parasitology, 116(3), 241-251. https://doi.org/10.1016/j.exppara.2007.01.008

Vancouver

Quiñones W, Peña P, Domingo-Sananes M, Cáceres A, Michels PAM, Avilan L et al. Leishmania mexicana: molecular cloning and characterization of enolase. Experimental Parasitology. 2007 Jul;116(3):241-251. https://doi.org/10.1016/j.exppara.2007.01.008

Author

Quiñones, Wilfredo ; Peña, Priscila ; Domingo-Sananes, Maria ; Cáceres, Ana ; Michels, Paul A. M. ; Avilan, Luisana ; Concepción, Juan Luis. / Leishmania mexicana : molecular cloning and characterization of enolase. In: Experimental Parasitology. 2007 ; Vol. 116, No. 3. pp. 241-251.

Bibtex

@article{eb27f8b063b546099af4baf61ad6724b,
title = "Leishmania mexicana: molecular cloning and characterization of enolase",
abstract = "The gene of Leishmania mexicana enolase was cloned and overexpressed in Escherichia coli as an active enzyme; the protein was biochemically analyzed. This enolase shares with enolases from other trypanosomatids the presence of three atypical residues, each with a reactive side group, near the active site, already described for the enzyme from Trypanosoma brucei. The natural enzyme was purified, using a three-step procedure, from a cytosolic fraction of L. mexicana promastigotes. The kinetic properties of the purified recombinant enzyme were similar to those of the natural enzyme. Both the recombinant and natural enzyme were inhibited by inorganic pyrophosphate. Subcellular localization analysis after differential centrifugation showed that the enzyme activity is only associated with the cytosolic fraction. However, an apparently inactive form of enolase was detected by Western blots in the microsomal fraction. Digitonin treatment of parasites and immunofluorescence studies with permeabilized and non-permeabilized parasites showed that enolase is also associated with membranes and it was found at the external face of the plasma membrane.",
keywords = "Amino Acid Sequence, Animals, Antibodies, Protozoan/immunology, Base Sequence, Blotting, Western, Cell Membrane Permeability/drug effects, Cloning, Molecular, Digitonin/pharmacology, Electrophoresis, Polyacrylamide Gel, Fluorescent Antibody Technique, Indicators and Reagents/pharmacology, Kinetics, Leishmania mexicana/enzymology, Mice, Molecular Sequence Data, Phosphopyruvate Hydratase/biosynthesis, Rabbits, Recombinant Proteins/biosynthesis, Sequence Alignment",
author = "Wilfredo Qui{\~n}ones and Priscila Pe{\~n}a and Maria Domingo-Sananes and Ana C{\'a}ceres and Michels, {Paul A. M.} and Luisana Avilan and Concepci{\'o}n, {Juan Luis}",
year = "2007",
month = jul,
doi = "10.1016/j.exppara.2007.01.008",
language = "English",
volume = "116",
pages = "241--251",
journal = "Experimental Parasitology",
issn = "0014-4894",
publisher = "Elsevier",
number = "3",

}

RIS

TY - JOUR

T1 - Leishmania mexicana

T2 - molecular cloning and characterization of enolase

AU - Quiñones, Wilfredo

AU - Peña, Priscila

AU - Domingo-Sananes, Maria

AU - Cáceres, Ana

AU - Michels, Paul A. M.

AU - Avilan, Luisana

AU - Concepción, Juan Luis

PY - 2007/7

Y1 - 2007/7

N2 - The gene of Leishmania mexicana enolase was cloned and overexpressed in Escherichia coli as an active enzyme; the protein was biochemically analyzed. This enolase shares with enolases from other trypanosomatids the presence of three atypical residues, each with a reactive side group, near the active site, already described for the enzyme from Trypanosoma brucei. The natural enzyme was purified, using a three-step procedure, from a cytosolic fraction of L. mexicana promastigotes. The kinetic properties of the purified recombinant enzyme were similar to those of the natural enzyme. Both the recombinant and natural enzyme were inhibited by inorganic pyrophosphate. Subcellular localization analysis after differential centrifugation showed that the enzyme activity is only associated with the cytosolic fraction. However, an apparently inactive form of enolase was detected by Western blots in the microsomal fraction. Digitonin treatment of parasites and immunofluorescence studies with permeabilized and non-permeabilized parasites showed that enolase is also associated with membranes and it was found at the external face of the plasma membrane.

AB - The gene of Leishmania mexicana enolase was cloned and overexpressed in Escherichia coli as an active enzyme; the protein was biochemically analyzed. This enolase shares with enolases from other trypanosomatids the presence of three atypical residues, each with a reactive side group, near the active site, already described for the enzyme from Trypanosoma brucei. The natural enzyme was purified, using a three-step procedure, from a cytosolic fraction of L. mexicana promastigotes. The kinetic properties of the purified recombinant enzyme were similar to those of the natural enzyme. Both the recombinant and natural enzyme were inhibited by inorganic pyrophosphate. Subcellular localization analysis after differential centrifugation showed that the enzyme activity is only associated with the cytosolic fraction. However, an apparently inactive form of enolase was detected by Western blots in the microsomal fraction. Digitonin treatment of parasites and immunofluorescence studies with permeabilized and non-permeabilized parasites showed that enolase is also associated with membranes and it was found at the external face of the plasma membrane.

KW - Amino Acid Sequence

KW - Animals

KW - Antibodies, Protozoan/immunology

KW - Base Sequence

KW - Blotting, Western

KW - Cell Membrane Permeability/drug effects

KW - Cloning, Molecular

KW - Digitonin/pharmacology

KW - Electrophoresis, Polyacrylamide Gel

KW - Fluorescent Antibody Technique

KW - Indicators and Reagents/pharmacology

KW - Kinetics

KW - Leishmania mexicana/enzymology

KW - Mice

KW - Molecular Sequence Data

KW - Phosphopyruvate Hydratase/biosynthesis

KW - Rabbits

KW - Recombinant Proteins/biosynthesis

KW - Sequence Alignment

UR - https://www.sciencedirect.com/science/article/pii/S0014489407000288?via%3Dihub

U2 - 10.1016/j.exppara.2007.01.008

DO - 10.1016/j.exppara.2007.01.008

M3 - Article

C2 - 17382932

VL - 116

SP - 241

EP - 251

JO - Experimental Parasitology

JF - Experimental Parasitology

SN - 0014-4894

IS - 3

ER -

ID: 24736434