Asparagine requirement in Plasmodium berghei as a target to prevent malaria transmission and liver infections

Viswanathan A Nagaraj,Pradeep A Subramani,Vinayagam Sathishkumar,Rajeev R Pandey,Susanta K Ghosh,Manjunatha C Shetty,Govindarajan Padmanaban,Dhanunjay Mukhi

doi:10.1038/ncomms9775

Abstract

The proteins of Plasmodium, the malaria parasite, are strikingly rich in asparagine. Plasmodium depends primarily on host haemoglobin degradation for amino acids and has a rudimentary pathway for amino acid biosynthesis, but retains a gene encoding asparagine synthetase (AS). Here we show that deletion of AS in Plasmodium berghei (Pb) delays the asexual- and liver-stage development with substantial reduction in the formation of ookinetes, oocysts and sporozoites in mosquitoes. In the absence of asparagine synthesis, extracellular asparagine supports suboptimal survival of PbAS knockout (KO) parasites. Depletion of blood asparagine levels by treating PbASKO-infected mice with asparaginase completely prevents the development of liver stages, exflagellation of male gametocytes and the subsequent formation of sexual stages. In vivo supplementation of asparagine in mice restores the exflagellation of PbASKO parasites. Thus, the parasite life cycle has an absolute requirement for asparagine, which we propose could be targeted to prevent malaria transmission and liver infections.

Highlights

The proteins of Plasmodium, the malaria parasite, are strikingly rich in asparagine
Asparagine biosynthesis is catalysed by two evolutionarily independent families of enzymes—asparagine synthetase (AS)-A and AS-B29,30—of which AS-A catalyses the amidation of aspartate using ammonia as nitrogen source, whereas AS-B can utilize both glutamine and ammonia though glutamine is preferred under physiological conditions (Fig. 1a)
While the presence of asparagine repeats in low complexity regions exists as a hallmark of P. falciparum proteins[34,35], asparagine serves as one of the most abundant amino acids in proteins of other Plasmodium species as well[18]

Summary

Introduction

The proteins of Plasmodium, the malaria parasite, are strikingly rich in asparagine. Plasmodium depends primarily on host haemoglobin degradation for amino acids and has a rudimentary pathway for amino acid biosynthesis, but retains a gene encoding asparagine synthetase (AS). The parasite life cycle has an absolute requirement for asparagine, which we propose could be targeted to prevent malaria transmission and liver infections. Auxotrophic for most of its amino acids, the parasite genome encodes a few enzymes synthesizing glycine, proline, glutamate, glutamine, aspartate and asparagine, the functional significance of which remains unknown[15,16] Of these amino acids, asparagine plays a pivotal role in the parasite life cycle by serving as one of the most abundant amino acids in Plasmodium proteins[18]. We use P. berghei as an in vivo rodent parasite model to address the significance of asparagine requirement in the entire life cycle of malaria parasites by performing targeted deletion of endogenous AS and depleting the extracellular asparagine by asparaginase treatment. Our study substantiates the therapeutic potential of amino acid requirements for targeting the multiple stages of malaria parasite life cycle

Methods

Results

Conclusion