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Fike, K. R.

Publications and source records attributed to Fike, K. R..

2 recordsLinked to original sources

Development of semisynthetic blasticidin S analogs with potent and fast-killing anti-malarial activity

Protein synthesis represents an attractive target space for the development of anti-malarials with novel modes of action. Natural-product inhibitors of the eukaryotic 80S ribosome can have potent anti-malarial activity but are often poorly selective due to mammalian cytotoxicity. Blasticidin S (BlaS) is a microbially-produced natural product that broadly inhibits prokaryotic and eukaryotic protein synthesis by binding to the ribosomal peptidyl transferase center. In this study, we explored the potential for improving the anti-malarial potency and selectivity of the blasticidin S scaffold with semi-synthetic analogs that are modified at the C6 and C4 sites. The two best analogs were two orders of magnitude more potent than BlaS against Plasmodium falciparum drug-sensitive and -resistant lines while displaying low cytotoxicity towards mammalian cells. These analogs exhibited improved kinetics of inhibition of protein synthesis in cultured parasites and blocked the development of asexual stages expressing the plasmodial surface anion channel, a transporter required for nutrient acquisition and BlaS uptake. They also exhibited a dramatically improved speed of killing over BlaS. Molecular docking analysis revealed that these analogs are able to form more interactions with the P. falciparum ribosomal peptidyl transferase center than is BlaS, which is consistent with their increased potency. Together, these studies demonstrate the feasibility of generating BlaS analogs with potent anti-malarial activity and provide a roadmap for further development.

microbiology↗

Defining the minimal enzymatic requirements for fatty acid scavenging from lysophosphatidylcholine by erythrocytic Plasmodium falciparum

Host-derived lysophosphatidylcholine (LPC) is a significant source of choline and fatty acids for the intraerythrocytic malaria parasite Plasmodium falciparum. Two lysophospholipases play a dominant role in LPC catabolism: exported lipase 2 (XL2) and exported lipase homolog 4 (XLH4). Loss of these two enzymes greatly reduces, but does not abrogate, the parasites ability to utilize LPC as a source of fatty acids. In this study, we identify a third enzyme, termed "prodrug activation and resistance esterase" (PARE), that mediates low levels of LPC hydrolysis. Loss of PARE alone had no effect on the parasites ability to scavenge fatty acids from LPC. However, when combined with the loss of XL2 and XLH4, knockdown of PARE impacted the parasites ability to scavenge both choline and fatty acids from LPC. Furthermore, PARE/XL2/XLH4-deficient parasites were unable to complete a replication cycle when cultured in defined media with LPC as the sole source of exogenous fatty acids. We show that PARE is a membrane-associated enzyme with a substantial presence at the parasite periphery and propose a model whereby PARE catalyzes the hydrolysis of inwardly-diffusing LPC. Our findings reveal that asexual P. falciparum is dependent on parasite-encoded enzymes for LPC catabolism and rule out host erythrocyte enzymes as a physiologically-relevant source of lysophospholipase activity.

microbiology↗