scholarly journals Characterization of Plasmodium falciparum Pantothenate Kinase and Identification of Its Inhibitors From Natural Products

2021 ◽  
Vol 11 ◽  
Author(s):  
Arif Nurkanto ◽  
Ghulam Jeelani ◽  
Herbert J. Santos ◽  
Yulia Rahmawati ◽  
Mihoko Mori ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Natural Products ◽  
Biosynthetic Pathway ◽  
Essential Role ◽  
Screening System ◽  
Metabolic Reactions ◽  
First Time ◽  
Rate Limiting ◽  
Primary Substrate

Coenzyme A (CoA) is a well-known cofactor that plays an essential role in many metabolic reactions in all organisms. In Plasmodium falciparum, the most deadly among Plasmodium species that cause malaria, CoA and its biosynthetic pathway have been proven to be indispensable. The first and rate-limiting reaction in the CoA biosynthetic pathway is catalyzed by two putative pantothenate kinases (PfPanK1 and 2) in this parasite. Here we produced, purified, and biochemically characterized recombinant PfPanK1 for the first time. PfPanK1 showed activity using pantetheine besides pantothenate, as the primary substrate, indicating that CoA biosynthesis in the blood stage of P. falciparum can bypass pantothenate. We further developed a robust and reliable screening system to identify inhibitors using recombinant PfPanK1 and identified four PfPanK inhibitors from natural compounds.

scholarly journals Biosynthesis of anandamide and N-palmitoylethanolamine by sequential actions of phospholipase A2 and lysophospholipase D

2004 ◽  
Vol 380 (3) ◽  
pp. 749-756 ◽  
Author(s):  
Yong-Xin SUN ◽  
Kazuhito TSUBOI ◽  
Yasuo OKAMOTO ◽  
Takeharu TONAI ◽  
Makoto MURAKAMI ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Brain Homogenate ◽  
Wide Distribution ◽  
Rat Tissues ◽  
Lysophospholipase D ◽  
Pla2 Enzyme ◽  
First Time ◽  
Hydrolysis Of ◽  
The Brain

Anandamide (an endocannabinoid) and other bioactive long-chain NAEs (N-acylethanolamines) are formed by direct release from N-acyl-PE (N-acyl-phosphatidylethanolamine) by a PLD (phospholipase D). However, the possible presence of a two-step pathway from N-acyl-PE has also been suggested previously, which comprises (1) the hydrolysis of N-acyl-PE to N-acyl-lysoPE by PLA1/PLA2 enzyme(s) and (2) the release of NAEs from N-acyllysoPE by lysoPLD (lysophospholipase D) enzyme(s). In the present study we report for the first time the characterization of enzymes responsible for this pathway. The PLA1/PLA2 activity for N-palmitoyl-PE was found in various rat tissues, with the highest activity in the stomach. This stomach enzyme was identified as group IB sPLA2 (secretory PLA2), and its product was determined as N-acyl-1-acyl-lysoPE. Recombinant group IB, IIA and V of sPLA2s were also active with N-palmitoyl-PE, whereas group X sPLA2 and cytosolic PLA2α were inactive. In addition, we found wide distribution of lysoPLD activity generating N-palmitoylethanolamine from N-palmitoyl-lysoPE in rat tissues, with higher activities in the brain and testis. Based on several lines of enzymological evidence, the lysoPLD enzyme could be distinct from the known N-acyl-PE-hydrolysing PLD. sPLA2-IB dose dependently enhanced the production of N-palmitoylethanolamine from N-palmitoyl-PE in the brain homogenate showing the lysoPLD activity. N-Arachidonoyl-PE and N-arachidonoyl-lysoPE as anandamide precursors were also good substrates of sPLA2-IB and the lysoPLD respectively. These results suggest that the sequential actions of PLA2 and lysoPLD may constitute another biosynthetic pathway for NAEs, including anandamide.

scholarly journals Antileishmanial Effect of 3-Aminooxy-1-Aminopropane Is Due to Polyamine Depletion

2006 ◽  
Vol 51 (2) ◽  
pp. 528-534 ◽  
Author(s):  
Sushma Singh ◽  
Angana Mukherjee ◽  
Alex R. Khomutov ◽  
Lo Persson ◽  
Olle Heby ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Leishmania Donovani ◽  
Clinical Isolates ◽  
Growth And Survival ◽  
Antiproliferative Effects ◽  
Cell Growth And Differentiation ◽  
First Time ◽  
Rate Limiting ◽  
Sodium Antimony Gluconate

ABSTRACT The polyamines putrescine, spermidine, and spermine are organic cations that are required for cell growth and differentiation. Ornithine decarboxylase (ODC), the first and rate-limiting enzyme in the polyamine biosynthetic pathway, catalyzes the conversion of ornithine to putrescine. As the polyamine biosynthetic pathway is essential for the growth and survival of Leishmania donovani, the causative agent of visceral leishmaniasis, inhibition of the pathway is an important leishmaniacidal strategy. In the present study, we examined for the first time the effects of 3-aminooxy-1-aminopropane (APA), an ODC inhibitor, on the growth of L. donovani. APA inhibited the growth of both promastigotes in vitro and amastigotes in the macrophage model, with the 50% inhibitory concentrations being 42 and 5 μM, respectively. However, concentrations of APA up to 200 μM did not affect the viability of macrophages. The effects of APA were completely abolished by the addition of putrescine or spermidine. APA induced a significant decrease in ODC activity and putrescine, spermidine, and trypanothione levels in L. donovani promastigotes. Parasites were transfected with an episomal ODC construct, and these ODC overexpressers exhibited significant resistance to APA and were concomitantly resistant to sodium antimony gluconate (Pentostam), indicating a role for ODC overexpression in antimonial drug resistance. Clinical isolates with sodium antimony gluconate resistance were also found to overexpress ODC and to have significant increases in putrescine and spermidine levels. However, no increase in trypanothione levels was observed. The ODC overexpression in these clinical isolates alleviated the antiproliferative effects of APA. Collectively, our results demonstrate that APA is a potent inhibitor of L. donovani growth and that its leishmaniacidal effect is due to inhibition of ODC.

Tropical Marine Algae. IX. A New Sesquiterpenoid Metabolite From the Red Alga Laurencia marianensis

1993 ◽  
Vol 46 (6) ◽  
pp. 933 ◽  
Author(s):  
R Denys ◽  
JC Coll ◽  
BF Bowden
Keyword(s):  
Natural Products ◽  
Marine Algae ◽  
Red Alga ◽  
Natural Products Chemistry ◽  
First Time

An investigation of the natural products chemistry of the red alga Laurencia marianensis Yamada, a species whose chemistry has not previously been described in the literature, yielded the new metabolite 1-[(3′S*,3a′lR*,4′R*,7′S*,7a′S*)-7′-bromo-7a′-methyl-3′-(1′-methylethyl)octahydro-1′H-inden-4′-yl] ethanone (1) and the known metabolites deoxyprepacifenol (2) and pacifenol (3). The full n.m.r. characterization of (2) and (3) is reported for the first time.

scholarly journals Characterization of the CDP-2-Glycerol Biosynthetic Pathway in Streptococcus pneumoniae

2010 ◽  
Vol 192 (20) ◽  
pp. 5506-5514 ◽  
Author(s):  
Quan Wang ◽  
Yanli Xu ◽  
Andrei V. Perepelov ◽  
Wei Xiong ◽  
Dongmei Wei ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Streptococcus Pneumoniae ◽  
Biosynthetic Pathway ◽  
Resonance Spectroscopy ◽  
Ionization Mass ◽  
Enzyme Substrate ◽  
Effects Of Temperature ◽  
First Time ◽  
Novel Products

ABSTRACT Capsule polysaccharide (CPS) plays an important role in the virulence of Streptococcus pneumoniae and is usually used as the pneumococcal vaccine target. Glycerol-2-phosphate is found in the CPS of S. pneumoniae types 15A and 23F and is rarely found in the polysaccharides of other bacteria. The biosynthetic pathway of the nucleotide-activated form of glycerol-2-phosphate (NDP-2-glycerol) has never been identified. In this study, three genes (gtp1, gtp2, and gtp3) from S. pneumoniae 23F that have been proposed to be involved in the synthesis of NDP-2-glycerol were cloned and the enzyme products were expressed, purified, and assayed for their respective activities. Capillary electrophoresis was used to detect novel products from the enzyme-substrate reactions, and the structure of the product was elucidated using electrospray ionization mass spectrometry and nuclear magnetic resonance spectroscopy. Gtp1 was identified as a reductase that catalyzes the conversion of 1,3-dihydroxyacetone to glycerol, Gtp3 was identified as a glycerol-2-phosphotransferase that catalyzes the conversion of glycerol to glycerol-2-phosphate, and Gtp2 was identified as a cytidylyltransferase that transfers CTP to glycerol-2-phosphate to form CDP-2-glycerol as the final product. The kinetic parameters of Gtp1 and Gtp2 were characterized in depth, and the effects of temperature, pH, and cations on these two enzymes were analyzed. This is the first time that the biosynthetic pathway of CDP-2-glycerol has been identified biochemically; this pathway provides a method to enzymatically synthesize this compound.

Syntheses of Cytosporones A, C, J, K, and N, Metabolites from Medicinal Fungi

2015 ◽  
Vol 68 (10) ◽  
pp. 1583 ◽  
Author(s):  
Andrew M. Beekman ◽  
Russell A. Barrow
Keyword(s):  
Natural Products ◽  
Selective Reduction ◽  
Complete Characterization ◽  
Phenyl Acetate ◽  
Human Pathogens ◽  
Medicinal Fungi ◽  
C And N ◽  
Key Steps ◽  
First Time

The syntheses of the fungal metabolites cytosporones A, (±)-C, and N are reported. And the syntheses of cytosporones J and K are described for the first time. The preparation of racemic cytosporone J and racemic cytosporone K, natural products containing the rare 3-isochromanone substructure, was achieved in 8 linear steps with an overall yield of 45 % and 7 linear steps in 46 % yield, respectively, resulting in the complete characterization of these compounds for the first time. The key steps included a recently described homologation of benzoic acid to the analogous phenyl acetate using Birch reductive alkylation conditions, acylation of the appropriate phenyl acetate derivative, and a selective reduction and spontaneous biomimetic lactonization to yield the 3-isochromanone skeleton. The synthesized natural products were evaluated for their biological activity against several clinical strains of human pathogens with all compounds displaying weak antimicrobial activity.

scholarly journals Characterization of the Endogenous DAF-12 Ligand and Its Use as an Anthelmintic Agent in Strongyloides stercoralis

2021 ◽  
Author(s):  
Zhu Wang ◽  
Mi Cheong Cheong ◽  
Jet Tsien ◽  
Heping Deng ◽  
Tian Qin ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Strongyloides Stercoralis ◽  
Preclinical Model ◽  
Loss Of Function ◽  
Drug Of Choice ◽  
Lethal Infection ◽  
Parasite Reproduction ◽  
Rate Limiting

ABSTRACTA prevalent feature of Strongyloides stercoralis is a life-long and potentially lethal infection that is due to the nematode parasite’s ability to autoinfect and, thereby, self-replicate within its host. Here, we investigated the role of the parasite’s nuclear receptor, Ss-DAF-12, in governing infection. We identified Δ7-DA as the endogenous Ss-DAF-12 ligand and elucidated the hormone’s biosynthetic pathway. Genetic loss of function of the ligand’s rate-limiting enzyme demonstrated that Δ7-DA synthesis is necessary for parasite reproduction, whereas its absence is required for development of infectious larvae. Availability of the ligand permits Ss-DAF-12 to function as an on/off switch governing autoinfection, making it vulnerable to therapeutic intervention. In a preclinical model of hyperinfection, pharmacologic activation of DAF-12 suppressed autoinfection and markedly reduced lethality. Moreover, when Δ7-DA was administered with ivermectin, the current but limited drug of choice for treating strongyloidiasis, the combinatorial effects of the two drugs resulted in a near cure of the disease.

scholarly journals Glucose-6-phosphate dehydrogenase–6-phosphogluconolactonase: a unique bifunctional enzyme from Plasmodium falciparum

2011 ◽  
Vol 436 (3) ◽  
pp. 641-650 ◽  
Author(s):  
Esther Jortzik ◽  
Boniface M. Mailu ◽  
Janina Preuss ◽  
Marina Fischer ◽  
Lars Bode ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Pentose Phosphate Pathway ◽  
Drug Target ◽  
Pentose Phosphate ◽  
Bifunctional Enzyme ◽  
Functional Differences ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
First Time ◽  
Human Rbcs

The survival of malaria parasites in human RBCs (red blood cells) depends on the pentose phosphate pathway, both in Plasmodium falciparum and its human host. G6PD (glucose-6-phosphate dehydrogenase) deficiency, the most common human enzyme deficiency, leads to a lack of NADPH in erythrocytes, and protects from malaria. In P. falciparum, G6PD is combined with the second enzyme of the pentose phosphate pathway to create a unique bifunctional enzyme named GluPho (glucose-6-phosphate dehydrogenase–6-phosphogluconolactonase). In the present paper, we report for the first time the cloning, heterologous overexpression, purification and kinetic characterization of both enzymatic activities of full-length PfGluPho (P. falciparum GluPho), and demonstrate striking structural and functional differences with the human enzymes. Detailed kinetic analyses indicate that PfGluPho functions on the basis of a rapid equilibrium random Bi Bi mechanism, where the binding of the second substrate depends on the first substrate. We furthermore show that PfGluPho is inhibited by S-glutathionylation. The availability of recombinant PfGluPho and the major differences to hG6PD (human G6PD) facilitate studies on PfGluPho as an excellent drug target candidate in the search for new antimalarial drugs.

scholarly journals Cooperative Involvement of Glycosyltransferases in the Transfer of Amino Sugars during the Biosynthesis of the Macrolactam Sipanmycin byStreptomycessp. Strain CS149

2018 ◽  
Vol 84 (18) ◽  
Author(s):  
Mónica G. Malmierca ◽  
Ignacio Pérez-Victoria ◽  
Jesús Martín ◽  
Fernando Reyes ◽  
Carmen Méndez ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Hydroxyl Group ◽  
Multiresistant Pathogens ◽  
Content Type ◽  
P450 Monooxygenases ◽  
Deoxy Sugar ◽  
Leaf Cutting ◽  
First Time

ABSTRACTMacrolactams comprise a family of natural compounds with important bioactivities, such as antibiotic, antifungal, and antiproliferative activities. Sipanmycins A and B are two novel members of this family, with two sugar moieties attached to the aglycon. In the related macrolactam vicenistatin, the sugar moiety has been proven to be essential for cytotoxicity. In this work, the gene cluster responsible for the biosynthesis of sipanmycins (sipcluster) inStreptomycessp. strain CS149 is described and the steps involved in the glycosylation of the final compounds unraveled. Also, the cooperation of two different glycosyltransferases in each glycosylation step is demonstrated. Additionally, the essential role of SipO2 as an auxiliary protein in the incorporation of the second deoxy sugar is addressed. In light of the results obtained by the generation of mutant strains andin silicocharacterization of thesipcluster, a biosynthetic pathway for sipanmycins and the two deoxy sugars attached is proposed. Finally, the importance of the hydroxyl group at C-10 of the macrolactam ring and the sugar moieties for cytotoxicity and antibiotic activity of sipanmycins is shown.IMPORTANCEThe rapid emergence of infectious diseases and multiresistant pathogens has increased the necessity for new bioactive compounds; thus, novel strategies have to be developed to find them. Actinomycetes isolated in symbiosis with insects have attracted attention in recent years as producers of metabolites with important bioactivities. Sipanmycins are glycosylated macrolactams produced byStreptomycessp. CS149, isolated from leaf-cutting ants, and show potent cytotoxic activity. Here, we characterize thesipcluster and propose a biosynthetic pathway for sipanmycins. As far as we know, it is the first time that the cooperation between two different glycosyltransferases is demonstrated to be strictly necessary for the incorporation of the same sugar. Also, a third protein with homology to P450 monooxygenases, SipO2, is shown to be essential in the second glycosylation step, forming a complex with the glycosyltransferase pair SipS9-SipS14.

scholarly journals Structural characterization of three noncanonical NTF2-like superfamily proteins: implications for polyketide biosynthesis

2020 ◽  
Vol 76 (8) ◽  
pp. 372-383
Author(s):  
Nemanja Vuksanovic ◽  
Xuechen Zhu ◽  
Dante A. Serrano ◽  
Vilja Siitonen ◽  
Mikko Metsä-Ketelä ◽  
...  
Keyword(s):  
Natural Products ◽  
Structural Characterization ◽  
Unknown Function ◽  
Biosynthetic Pathway ◽  
Streptomyces Coelicolor ◽  
Biosynthetic Pathways ◽  
Polyketide Biosynthesis ◽  
X Ray ◽  
Bona Fide

Proteins belonging to the NTF2-like superfamily are present in the biosynthetic pathways of numerous polyketide natural products, such as anthracyclins and benzoisochromanequinones. Some have been found to be bona fide polyketide cyclases, but many of them have roles that are currently unknown. Here, the X-ray crystal structures of three NTF2-like proteins of unknown function are reported: those of ActVI-ORFA from Streptomyces coelicolor A3(2) and its homologs Caci_6494, a protein from an uncharacterized biosynthetic cluster in Catenulispora acidiphila, and Aln2 from Streptomyces sp. CM020, a protein in the biosynthetic pathway of alnumycin. The presence of a solvent-accessible cavity and the conservation of the His/Asp dyad that is characteristic of many polyketide cyclases suggest a potential enzymatic role for these enzymes in polyketide biosynthesis.

Investigation of Antiplasmodial Effects of Terpenoid Compounds Isolated from Myrrh

Planta Medica ◽  
2020 ◽  
Vol 86 (09) ◽  
pp. 643-654 ◽  
Author(s):  
Hippolyt L. Greve ◽  
Marcel Kaiser ◽  
Thomas J. Schmidt
Keyword(s):  
Plasmodium Falciparum ◽  
In Vitro Activity ◽  
Isolation And Characterization ◽  
Spectroscopic Analyses ◽  
Ic50 Value ◽  
Ic50 Values ◽  
O 18 ◽  
First Time

AbstractAs part of our ongoing search for antiprotozoal natural products from plants, we examined different resins from the Burseraceae family. The dichloromethane extract obtained from myrrh, the oleo-gum-resin of Commiphora species, showed promising in vitro activity against Plasmodium falciparum with an IC50 value of 1 µg/mL. Bioactivity-guided fractionation led to the isolation and characterization of 18 sesquiterpenoids, namely, β-elemene (1), elemyl acetate (2), curzerenone (3), 8-hydroxyisogermafurenolide (4), 2-methoxyisogermafurenolide (5), 8-epi-2-methoxyisogermafurenolide (6), furanodienone (7), 1(10)Z,4Z-furanodien-6-one (8), rel-2R-methyl-5S-acetoxy-4R-furanogermacr-1(10)Z-en-6-one (9), (1(10)E)-2-methoxy-8,12-epoxygermacra-1(10),7,11-trien-6-one (10), 2R-methoxyfuranodiene (11), 2-acetyloxyglechomanolide (12), 8-epi-2-acetyloxyglechomanolide (13), (1R,2R,4S)-1,2-epoxyfuranogermacr-10(15)-en-6-one (14), hydroxylindestrenolide (15), isohydroxylindestrenolide (16), myrrhone (17), and myrrhterpenoid O (18). Moreover, nine (nor-)triterpenoids were isolated: mansumbinol (19), mansumbinol epoxide (20), mansumbinone (21), mansumbin-13(17)-en-3,16-dione (22), 3,4-seco-mansumbinoic acid (23), rel-20S-hydroxy-dammar-24-en-3,16-dione (24), rel-(16S,20S)-dihydroxydammar-24-en-3-one (25), cycloart-24-en-1α,2α,3β-triol (26), and 3β-isovaleroyloxycycloart-24-en-1α,2α-diol (27). All compounds were identified by MS and NMR spectroscopic analyses. To the best of our knowledge, compounds 5, 6, 12, 13, 16, 18, and 20 are reported for the first time. All isolated compounds were tested in vitro for activity against P. falciparum and cytotoxicity. The sesquiterpene 7 and the triterpene 25 were the most active compounds found in this study with IC50 values of 7.4 and 2.8 µM, respectively.

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