Cloning of a pyruvate phosphate dikinase from Trypanosoma cruzi

TY - JOUR

T1 - Cloning of a pyruvate phosphate dikinase from Trypanosoma cruzi

AU - Maldonado, Rosa A.

AU - Fairlamb, Alan H.

N1 - Funding Information: This work was supported by an International Travelling Fellowship to RAM from the Wellcome Trust. We would like to thank Dr E Tetaud for comments, Alex Shuttlekopf for assistance in preparation of the structural model and to Dr. F. Bringaud and Prof. T. Baltz for the PPDK monoclonal antibody.

PY - 2001/2

Y1 - 2001/2

N2 - We have cloned and characterised a gene that encodes a putative pyruvate phosphate dikinase (PPDK) from Trypanosoma cruzi, an enzyme that catalyses the reversible conversion of phosphoenolpyruvate to pyruvate. PPDK is absent in mammalian cells, but has been found in a wide variety of other organisms, including plants and bacteria. In T. cruzi, two genes (PPDK1 and PPDK2) are present in a tandem array localised on a 1 Mbp chromosome. Northern and Western blot analyses indicates that PPDK is expressed as a 100-kDa protein in epimastigote, amastigote and trypomastigote forms. PPDK1 and PPDK2 encode an identical protein of 100.8 kDa with a C-terminal extension ending with the sequence AKL, a signal for glycosomal import. Both T. cruzi and T. brucei enzymes possess a 23-residue insertion, that is absent in other PPDKs. A three-dimensional alignment with the crystal structure of the enzyme from Clostridium symbiosum predicts that this insertion is located on the surface of the nucleotide-binding domain. Phylogenetic studies indicate that bacterial and protist PPDKs cluster as a separate group from those of plants. The evolutionary implications and possible role of this enzyme in T. cruzi is discussed.

AB - We have cloned and characterised a gene that encodes a putative pyruvate phosphate dikinase (PPDK) from Trypanosoma cruzi, an enzyme that catalyses the reversible conversion of phosphoenolpyruvate to pyruvate. PPDK is absent in mammalian cells, but has been found in a wide variety of other organisms, including plants and bacteria. In T. cruzi, two genes (PPDK1 and PPDK2) are present in a tandem array localised on a 1 Mbp chromosome. Northern and Western blot analyses indicates that PPDK is expressed as a 100-kDa protein in epimastigote, amastigote and trypomastigote forms. PPDK1 and PPDK2 encode an identical protein of 100.8 kDa with a C-terminal extension ending with the sequence AKL, a signal for glycosomal import. Both T. cruzi and T. brucei enzymes possess a 23-residue insertion, that is absent in other PPDKs. A three-dimensional alignment with the crystal structure of the enzyme from Clostridium symbiosum predicts that this insertion is located on the surface of the nucleotide-binding domain. Phylogenetic studies indicate that bacterial and protist PPDKs cluster as a separate group from those of plants. The evolutionary implications and possible role of this enzyme in T. cruzi is discussed.

KW - Gluconeogenesis

KW - Metabolism

KW - Pyruvate orthophosphate dikinase

KW - Trypanosoma

UR - https://www.scopus.com/pages/publications/0035115884

U2 - 10.1016/S0166-6851(00)00362-5

DO - 10.1016/S0166-6851(00)00362-5

M3 - Article

C2 - 11223125

AN - SCOPUS:0035115884

SN - 0166-6851

VL - 112

SP - 183

EP - 191

JO - Molecular and Biochemical Parasitology

JF - Molecular and Biochemical Parasitology

IS - 2

ER -