Search bioRxiv⌕ Search

Biology subjects

Grotli, M.

Publications and source records attributed to Grotli, M..

3 recordsLinked to original sources

Identification of potent and selective N-myristoyltransferase inhibitors of Plasmodium vivax liver stage hypnozoites and schizonts

New drugs targeting multiple stages of the malaria-causing parasite, Plasmodium, are needed to reduce and eliminate malaria worldwide. N-Myristoyltransferase (NMT) is an essential eukaryotic enzyme, and a validated chemically tractable drug target for malaria. Previous efforts have failed to target NMT owing to the low selectivity for the Plasmodium enzyme compared with human NMTs. Herein, we applied a structure-guided approach using previously reported NMT inhibitors as scaffolds to develop a new generation of Plasmodium vivax NMT (PvNMT) targeting compounds. We report a series of compounds with IC50 values in the nM range and an order of magnitude improved selectivity to Plasmodium NMT over human NMT (HsNMT). X-ray co-crystallization of PvNMT with a representative lead compound, 12b, supported the prevailing hypothesis that a conformational difference in a key tyrosine residue of PvNMT and HsNMT drives the selectivity between these enzymes. The compounds were triaged based on their selectivity for PvNMT. They significantly decreased P. falciparum blood-stage parasite load, with IC50 values ranging from 0.36 M to 1.25 M. The compounds exhibited a dose-dependent inhibition of P. vivax liver stage schizont and hypnozoite infection, consistently, in three different P. vivax isolates with IC50 values ranging from 2.2 M to 6 M and from 1.2 M to 12 M. Our data provide evidence that NMT inhibitors could be multistage antimalarials, targeting both dormant and developing liver stage parasites, which is essential for malaria elimination. One Sentence SummaryPotent and selective N-myristoyltransferase inhibitors of Plasmodium vivax hypnozoites and schizonts were synthesized and tested.

pharmacology and toxicology↗

Peroxiredoxin promotes longevity and H2O2-resistance in yeast through redox-modification of PKA

Peroxiredoxins are H2O2 scavenging enzymes that also carry H2O2 signaling and chaperone functions. In yeast, the major cytosolic peroxiredoxin, Tsa1 is required for both promoting resistance to H2O2 and extending lifespan upon caloric restriction. We show here that Tsa1 effects both these functions not by scavenging H2O2, but by repressing the nutrient signaling Ras-cAMP-PKA pathway at the level of the protein kinase A (PKA) enzyme. Tsa1 stimulates sulfenylation of cysteines in the PKA catalytic subunit by H2O2 and a significant proportion of the catalytic subunits are glutathionylated on two cysteine residues. Redox modification of the conserved Cys243 inhibits the phosphorylation of a conserved Thr241 in the kinase activation loop and enzyme activity, and preventing Thr241 phosphorylation can overcome the H2O2 sensitivity of Tsa1-deficient cells. Results support a model of aging where nutrient signaling pathways constitute hubs integrating information from multiple aging-related conduits, including a peroxiredoxin-dependent response to H2O2.

biochemistry↗

Discovery of functional alternatively spliced PKM transcripts in human cancers

The association of pyruvate kinase muscle type (PKM) with survival of cancer patients is controversial. Here, we focus on different transcripts of PKM and investigate the association between their mRNA expression and the clinical survival of the patients in 25 different cancers. We find that the transcript encoding PKM2, and three other functional transcripts are prognostic in multiple cancers. Our integrative analysis shows that the functions of these four transcripts are highly conservative in different cancers. Next, we validate the prognostic effect of these transcripts in an independent kidney renal clear-cell carcinoma (KIRC) cohort and identify a prognostic signature which could distinguish high- and low-risk KIRC patients. Finally, we reveal the functional role of alternatively spliced PKM transcripts in KIRC, and discover the protein products of different transcripts of PKM. Our analysis demonstrated that alternatively spliced transcripts of not only PKM but also other genes should be considered in cancer studies, since it may enable the discovery and targeting of the right protein product for development of the efficient treatment strategies.

systems biology↗