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Phosphofructokinase from Plasmodium berghei. Influence of Mg2+, ATP and Mg2(+)-complexed ATP.

ABSTRACT: The control enzyme phosphofructokinase is of regulatory significance in the metabolism of glucose by the malarial parasite Plasmodium berghei. (1) The enzyme was partially purified from erythrocytic stages of P. berghei by precipitation with poly(ethylene glycol) and chromatography on 2',5'-bisphosphoadenosine-Sepharose 4B. (2) Similarly to various other phosphofructokinases, the enzyme from P. berghei shows an allosteric behaviour. It is activated by fructose 6-phosphate and inhibited by ATP. (3) The effects of Mg2(+)-complexed ATP, free ATP and Mg2+ were studied by keeping constant the concentration of one of these and varying the concentrations of the other two. (4) The enzyme is shown to be allosterically inhibited by free ATP and by higher concentrations of Mg2+. Compared with phosphofructokinase of erythrocytes, inhibition by ATP is weaker by two orders of magnitude. Mg2(+)-complexed ATP has no effect on allosteric regulation. (5) The proposed kinetic model provides an adequate description of the data.

SUBMITTER: Buckwitz D

PROVIDER: S-EPMC1131295 | biostudies-other | 1990 Apr

REPOSITORIES: biostudies-other

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Project description:BackgroundThe native gut microbiota of Anopheles mosquitoes is known to play a key role in the physiological function of its host. Interestingly, this microbiota can also influence the development of Plasmodium in its host mosquitoes. In recent years, much interest has been shown in the employment of gut symbionts derived from vectors in the control of vector-borne disease transmission. In this study, the midgut microbial diversity has been characterized among laboratory-reared adult Anopheles stephensi mosquitoes, from the colony created by rearing progeny of wild-caught mosquitoes (obtained from three different locations in southern India) for multiple generations, using 16S ribosomal RNA (rRNA) gene sequencing approach. Further, the influence of native midgut microbiota of mosquitoes on the development of rodent malaria parasite Plasmodium berghei in its host has been studied.MethodsThe microbial diversity associated with the midgut of An. stephensi mosquitoes was studied by sequencing V3 region of 16S ribosomal RNA (rRNA) gene. The influence of native midgut microbiota of An. stephensi mosquitoes on the susceptibility of the mosquitoes to rodent malaria parasite P. berghei was studied by comparing the intensity and prevalence of P. berghei infection among the antibiotic treated and untreated cohorts of mosquitoes.ResultsThe analysis of bacterial diversity from the midguts of An. stephensi showed Proteobacteria as the most dominant population among the three laboratory-reared strains of An. stephensi studied. Major genera identified among these mosquito strains were Acinetobacter, Pseudomonas, Prevotella, Corynebacterium, Veillonella, and Bacillus. The mosquito infectivity studies carried out to determine the implication of total midgut microbiota on P. berghei infection showed that mosquitoes whose native microbiota cleared with antibiotics had increased susceptibility to P. berghei infection compared to the antibiotic untreated mosquitoes with its natural native microbiota.ConclusionsThe use of microbial symbiont to reduce the competence of vectors involved in disease transmission has gained much importance in recent years as an emerging alternative approach towards disease control. In this context, the present study was aimed to identify the midgut microbiota composition of An. stephensi, and its effect on the development of P. berghei. Interestingly, the analysis of midgut microbiota from An. stephensi revealed the presence of genus Veillonella in Anopheles species for the first time. Importantly, the study also revealed the negative influence of total midgut microbiota on the development of P. berghei in three laboratory strains of An. stephensi, emphasizing the importance of understanding the gut microbiota in malaria vectors, and its relationship with parasite development in designing strategies to control malaria transmission.

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Phosphofructokinase from Plasmodium berghei. Influence of Mg2+, ATP and Mg2(+)-complexed ATP.

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