Cloning, expression and biochemical characterization of mitochondrial and cytosolic malate dehydrogenase from Phytophthora infestans

Patricia E López-Calcagno; Johanna Moreno; Luis Cedeño; Luis Labrador; Juan L Concepción; Luisana Avilán

doi:10.1016/j.mycres.2009.02.012

Cloning, expression and biochemical characterization of mitochondrial and cytosolic malate dehydrogenase from Phytophthora infestans

Patricia E López-Calcagno, Johanna Moreno, Luis Cedeño, Luis Labrador, Juan L Concepción, Luisana Avilán^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

The genes of the mitochondrial and cytosolic malate dehydrogenase (mMDH and cMDH) of Phytophthora infestans were cloned and overexpressed in Escherichia coli as active enzymes. The catalytic properties of these proteins were determined: both enzymes have a similar specific activity. In addition, the natural mitochondrial isoenzyme was semi-purified from mycelia and its catalytic properties determined: the recombinant mitochondrial isoform behaved as the natural enzyme. A phylogenetic analysis indicated that mMDH, present in all stramenopiles studied, can be useful to study the relationships between these organisms. MDH with the conserved domain MDH_cytoplasmic_cytosolic is absent in some stramenopiles as well as in fungi. This enzyme seems to be less related within the stramenopile group. The Phytophthora cMDHs have an insertion of six amino acids that is also present in the stramenopile cMDHs studied, with the exception of Thalassiosira pseudonana cMDH, and is absent in other known eukaryotic cMDHs.

Original language	English
Pages (from-to)	771-81
Number of pages	11
Journal	Mycological Research
Volume	113
Issue number	Pt 6-7
Early online date	26 Feb 2009
DOIs	https://doi.org/10.1016/j.mycres.2009.02.012
Publication status	Published - Jun 2009

Keywords

Amino Acid Sequence
Cloning, Molecular
Cytosol/chemistry
Escherichia coli/genetics
Gene Expression
Kinetics
Malate Dehydrogenase/chemistry
Mitochondria/chemistry
Molecular Sequence Data
Oomycetes/chemistry
Phylogeny
Phytophthora infestans/chemistry
Sequence Alignment

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10.1016/j.mycres.2009.02.012

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@article{1ee2f3b351ad48f9a8d6dcf42e3afb70,

title = "Cloning, expression and biochemical characterization of mitochondrial and cytosolic malate dehydrogenase from Phytophthora infestans",

abstract = "The genes of the mitochondrial and cytosolic malate dehydrogenase (mMDH and cMDH) of Phytophthora infestans were cloned and overexpressed in Escherichia coli as active enzymes. The catalytic properties of these proteins were determined: both enzymes have a similar specific activity. In addition, the natural mitochondrial isoenzyme was semi-purified from mycelia and its catalytic properties determined: the recombinant mitochondrial isoform behaved as the natural enzyme. A phylogenetic analysis indicated that mMDH, present in all stramenopiles studied, can be useful to study the relationships between these organisms. MDH with the conserved domain MDH_cytoplasmic_cytosolic is absent in some stramenopiles as well as in fungi. This enzyme seems to be less related within the stramenopile group. The Phytophthora cMDHs have an insertion of six amino acids that is also present in the stramenopile cMDHs studied, with the exception of Thalassiosira pseudonana cMDH, and is absent in other known eukaryotic cMDHs.",

keywords = "Amino Acid Sequence, Cloning, Molecular, Cytosol/chemistry, Escherichia coli/genetics, Gene Expression, Kinetics, Malate Dehydrogenase/chemistry, Mitochondria/chemistry, Molecular Sequence Data, Oomycetes/chemistry, Phylogeny, Phytophthora infestans/chemistry, Sequence Alignment",

author = "L{\'o}pez-Calcagno, {Patricia E} and Johanna Moreno and Luis Cede{\~n}o and Luis Labrador and Concepci{\'o}n, {Juan L} and Luisana Avil{\'a}n",

year = "2009",

month = jun,

doi = "10.1016/j.mycres.2009.02.012",

language = "English",

volume = "113",

pages = "771--81",

journal = "Mycological Research",

issn = "0953-7562",

publisher = "Elsevier",

number = "Pt 6-7",

}

TY - JOUR

T1 - Cloning, expression and biochemical characterization of mitochondrial and cytosolic malate dehydrogenase from Phytophthora infestans

AU - López-Calcagno, Patricia E

AU - Moreno, Johanna

AU - Cedeño, Luis

AU - Labrador, Luis

AU - Concepción, Juan L

AU - Avilán, Luisana

PY - 2009/6

Y1 - 2009/6

N2 - The genes of the mitochondrial and cytosolic malate dehydrogenase (mMDH and cMDH) of Phytophthora infestans were cloned and overexpressed in Escherichia coli as active enzymes. The catalytic properties of these proteins were determined: both enzymes have a similar specific activity. In addition, the natural mitochondrial isoenzyme was semi-purified from mycelia and its catalytic properties determined: the recombinant mitochondrial isoform behaved as the natural enzyme. A phylogenetic analysis indicated that mMDH, present in all stramenopiles studied, can be useful to study the relationships between these organisms. MDH with the conserved domain MDH_cytoplasmic_cytosolic is absent in some stramenopiles as well as in fungi. This enzyme seems to be less related within the stramenopile group. The Phytophthora cMDHs have an insertion of six amino acids that is also present in the stramenopile cMDHs studied, with the exception of Thalassiosira pseudonana cMDH, and is absent in other known eukaryotic cMDHs.

AB - The genes of the mitochondrial and cytosolic malate dehydrogenase (mMDH and cMDH) of Phytophthora infestans were cloned and overexpressed in Escherichia coli as active enzymes. The catalytic properties of these proteins were determined: both enzymes have a similar specific activity. In addition, the natural mitochondrial isoenzyme was semi-purified from mycelia and its catalytic properties determined: the recombinant mitochondrial isoform behaved as the natural enzyme. A phylogenetic analysis indicated that mMDH, present in all stramenopiles studied, can be useful to study the relationships between these organisms. MDH with the conserved domain MDH_cytoplasmic_cytosolic is absent in some stramenopiles as well as in fungi. This enzyme seems to be less related within the stramenopile group. The Phytophthora cMDHs have an insertion of six amino acids that is also present in the stramenopile cMDHs studied, with the exception of Thalassiosira pseudonana cMDH, and is absent in other known eukaryotic cMDHs.

KW - Amino Acid Sequence

KW - Cloning, Molecular

KW - Cytosol/chemistry

KW - Escherichia coli/genetics

KW - Gene Expression

KW - Kinetics

KW - Malate Dehydrogenase/chemistry

KW - Mitochondria/chemistry

KW - Molecular Sequence Data

KW - Oomycetes/chemistry

KW - Phylogeny

KW - Phytophthora infestans/chemistry

KW - Sequence Alignment

UR - https://www.sciencedirect.com/science/article/pii/S0953756209000549?via%3Dihub

U2 - 10.1016/j.mycres.2009.02.012

DO - 10.1016/j.mycres.2009.02.012

M3 - Article

C2 - 19249364

SN - 0953-7562

VL - 113

SP - 771

EP - 781

JO - Mycological Research

JF - Mycological Research

IS - Pt 6-7

ER -

Cloning, expression and biochemical characterization of mitochondrial and cytosolic malate dehydrogenase from Phytophthora infestans

Abstract

Keywords

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