Orotate phosphoribosyltransferase and orotidine 5′-monophosphate decarboxylase exist as multienzyme complex in human malaria parasite Plasmodium falciparum

Krungkrai, Sudaratana R.; Prapunwattana, Phisit; Horii, Toshihiro; Krungkrai, Jerapan

doi:10.1016/j.bbrc.2004.04.124

Cited by 34 publications

(33 citation statements)

References 35 publications

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“…OMPD. Decarboxylation of orotidine-5=-monophosphate (OMP) to UMP was monitored directly at 285 nm (37). The 500-l reaction mixture contained 50 mM potassium phosphate buffer, pH 7.0, 2 mM DTT, and 0.1 mM EDTA.…”

Section: Methodsmentioning

confidence: 99%

Expression of Functional Plasmodium falciparum Enzymes Using a Wheat Germ Cell-Free System

Mudeppa

Rathod

2013

Eukaryot Cell

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One decade after the sequencing of the Plasmodium falciparum genome, 95% of malaria proteins in the genome cannot be expressed in traditional cell-based expression systems, and the targets of the best new leads for antimalarial drug discovery are either not known or not available in functional form. For a disease that kills up to 1 million people per year, routine expression of recombinant malaria proteins in functional form is needed both for the discovery of new therapeutics and for identification of targets of new drugs. We tested the general utility of cell-free systems for expressing malaria enzymes. Thirteen test enzyme sequences were reverse amplified from total RNA, cloned into a plant-like expression vector, and subjected to cell-free expression in a wheat germ system. Protein electrophoresis and autoradiography confirmed the synthesis of products of expected molecular masses. In rare problematic cases, truncated products were avoided by using synthetic genes carrying wheat codons. Scaled-up production generated 39 to 354 g of soluble protein per 10 mg of translation lysate. Compared to rare proteins where cell-based systems do produce functional proteins, the cell-free yields are comparable or better. All 13 test products were enzymatically active, without failure. This general path to produce functional malaria proteins should now allow the community to access new tools, such as biologically active protein arrays, and lead to the discovery of new chemical functions, structures, and inhibitors of previously inaccessible malaria gene products.

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Section: Methodsmentioning

confidence: 99%

Expression of Functional Plasmodium falciparum Enzymes Using a Wheat Germ Cell-Free System

Mudeppa

Rathod

2013

Eukaryot Cell

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“…In intra-erythrocytic stage of development, the malarial parasites require purines and pyrimidines for DNA/RNA synthesis and other metabolisms during this exponential growth and replication. The parasites, known as purine auxotroph, salvage the preformed purines from the mammalian host, but they have to synthesize pyrimidines de novo from HCO 3 -, adenine 5'-triphosphate, glutamine, aspartate and 5-phosphoribosyl-1-pyrophosphate [25][26][27][28][29][30][31][32][33]. These properties on both purine and pyrimidine biosynthesis are key differences between the parasite and human [34,35].…”

Section: Malarial Parasite Carbonic Anhydrasesmentioning

confidence: 99%

Malarial Parasite Carbonic Anhydrase and Its Inhibitors

Krungkrai¹,

Krungkrai²,

Supuran

2007

CTMC

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Plasmodium falciparum is responsible for the majority of life-threatening cases of human malaria. The global emergence of drug-resistant malarial parasites necessitates identification and characterization of novel drug targets. At present, carbonic anhydrase (CA) genes are identified in limited numbers of protozoa and helminthes parasites, however, they are demonstrated in at least 4 Plasmodium species. The CA gene of P. falciparum encodes an alpha-carbonic anhydrase enzyme possessing catalytic properties distinct of that of the human host enzymes. A small library of aromatic sulfonamides, most of which were Schiff's bases derived from sulfanilamide/homosulfanilamide/4-aminoethylbenzenesulfonamide and substituted-aromatic aldehydes, or ureido-substituted sulfonamides are good inhibitors of P. falciparum enzyme. The 4-(3,4-dichlorophenylureido-ethyl)-benzenesulfonamide is the most effective antimalarial activity against growth of P. falciparum in vitro. The nature of the groups substituting the aromatic-ureido- or aromatic-azomethine fragment of the molecule and the length of the parent sulfonamide (i.e., from sulfanilamide to 4-aminoethylbenzenesulfonamide) from which the Schiff's base obtained, are the critical parameters for the CA inhibitory activities of these aromatic sulfonamide derivatives, both against the malarial as well as human enzymes. Thus, the sulfonamide CA inhibitors may have the potential for the development of novel antimalarial drugs.

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“…It salvages the preformed purine bases/nucleosides from the human host and converts them to their mono-, di-and triphosphates. The parasite can only synthesize pyrimidines de novo from HCO 3 -, ATP, glutamine, aspartate, and 5-phosphoribosyl-1-pyrophosphate [43][44][45][46][47][48][49][50][51][52][53] . These unique properties on both purine and pyrimidine requirement of the parasite are key differences from the human host, in which both functional de novo and salvage pathways of the purine and pyrimidine synthesis exists [46,48,[54][55][56][57][58].…”

Section: Basic Life Cycle Genomics and Biochemistry Of Human Malariamentioning

confidence: 99%

“…Indeed, these sulfonamides have been assayed earlier for the inhibition of the human CA I, II, IV and IX isozymes, showing a great variation of potency and different affinities for the various human isozymes. Table 3 In vitro inhibition data of pfCA (KI, 毺 M), in vitro inhibition of growth of P. falciparum in cell cultures (IC 50 , 毺 M) and in vivo antimalarial activity in P. berghei infected mice (ID 50 , mg/kg) of sulfonamides Acetazolamide AZA and antimalarial drugs quinine, qinghausu and chloroquine, as standards. Compounds 10,16 and AZA are tested at 25.0, 10.0, 5.0, 2.5 mg/kg body weight for the in vivo study.…”

Section: Inhibition Of Malaria Parasite 毩 -Carbonic Anhydrase By Arommentioning

confidence: 99%

Malaria parasite carbonic anhydrase: inhibition of aromatic/heterocyclic sulfonamides and its therapeutic potential

Krungkrai¹,

Krungkrai²

2011

Asian Pacific Journal of Tropical Biomedicine

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Orotate phosphoribosyltransferase and orotidine 5′-monophosphate decarboxylase exist as multienzyme complex in human malaria parasite Plasmodium falciparum

Cited by 34 publications

References 35 publications

Expression of Functional Plasmodium falciparum Enzymes Using a Wheat Germ Cell-Free System

Expression of Functional Plasmodium falciparum Enzymes Using a Wheat Germ Cell-Free System

Malarial Parasite Carbonic Anhydrase and Its Inhibitors

Malaria parasite carbonic anhydrase: inhibition of aromatic/heterocyclic sulfonamides and its therapeutic potential

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