Analysis Biomolecular Interactions of the Inhibitor MMV007389 with PfFNT by BLI (CAT#: STEM-MB-0212-CJ)

Introduction

The emergence and spread of resistance by strains of P. falciparum to major antimalarial drugs has intensified the need to discover and develop drugs that kill the parasite by new mechanisms. The compound MMV007839 disrupts pH and kills the parasite's mechanism. Has been found to inhibit the Plasmodium falciparum nitrite formate transporter (PfFNT). PfFNT is a transporter on the parasite surface that is used to remove waste lactic acid from the parasite. The compound could not only inhibit the transp+E280ort of H+ -coupled lactic acid in parasite plasma membrane, but also inhibit the transport of lactic acid by PfFNT in Xenopus oocytes. In addition to disrupting pH, PfFNT inhibition can also cause lactic acid buildup in parasite-infected red blood cells and swelling of both the parasite and infected red blood cells. Exposing the parasite to MMV007839 for a long period of time produced a resistant parasite with a mutant form of PfFNT that was less sensitive to the compound.

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Principle Applications Procedure Materials Notes Related Products

Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Pharmacology

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.

Materials

• Equipment: Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules

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