3 years ago

Heterologous Expression, Purification, and Functional Analysis of Plasmodium falciparum Phosphatidylinositol 3′-Kinase

Heterologous Expression, Purification, and Functional Analysis of Plasmodium falciparum Phosphatidylinositol 3′-Kinase
Paul D. Roepe, Anna R. Sternberg, Craig J. Thomas, Bryce E. Riegel, Matthew R. Hassett
The Plasmodium falciparum malarial parasite genome appears to encode one and only one phosphatidylinositol 3′-kinase (PI3K), and sequence analysis suggests that the enzyme is a “class III”- or “Vps34”-type PI3K. PfVps34 has generated excitement as a possible druggable target and potentially a key target of artemisinin-based antimalarials. In this study, we optimize the PfVps34 gene for heterologous expression in yeast, purify the protein to homogeneity, use a recently validated quantitative assay for phosphatidylinositol 3′-phosphate production from phosphatidylinositol (Hassett et al., companion paper; DOI 10.1021/acs.biochem.7b00416) to quantify activity and drug inhibition of that activity, and investigate the importance of key residues in the enzyme’s catalytic and “N-lobe” domains. Data suggest that PfVps34 is indeed inhibited by artemisinin and related drugs but only under conditions that cleave the drugs’ endoperoxide bridge to generate reactive alkylating agents.

Publisher URL: http://dx.doi.org/10.1021/acs.biochem.7b00416

DOI: 10.1021/acs.biochem.7b00416

You might also like
Discover & Discuss Important Research

Keeping up-to-date with research can feel impossible, with papers being published faster than you'll ever be able to read them. That's where Researcher comes in: we're simplifying discovery and making important discussions happen. With over 19,000 sources, including peer-reviewed journals, preprints, blogs, universities, podcasts and Live events across 10 research areas, you'll never miss what's important to you. It's like social media, but better. Oh, and we should mention - it's free.

  • Download from Google Play
  • Download from App Store
  • Download from AppInChina

Researcher displays publicly available abstracts and doesn’t host any full article content. If the content is open access, we will direct clicks from the abstracts to the publisher website and display the PDF copy on our platform. Clicks to view the full text will be directed to the publisher website, where only users with subscriptions or access through their institution are able to view the full article.