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New Reference – PiColorLock™

PiColorLock™ has been published in Nature!

Tuberculosis (TB) is a major health concern and worldwide emergence of resistant forms of Mycobacterium tuberculosis (Mtb) has further aggravated the situation. As most of the available clinical drugs have single targets in the pathogens, these drugs become ineffective over time due to the development of bacterial resistance against them. The development of new drugs against TB should address the pathogen’s resistance mechanisms rather than the discovery of a novel inactivation target. Furthermore, the development of new anti-TB drugs should be focused on a multi-targeted therapy, where a single drug can target multiple enzymes in key metabolic pathways to reduce the chances of the pathogen developing drug-resistance.

In this paper the authors focus on the Mur family, enzymes that catalyse the biosynthesis of UDP-n-acetylmuramyl-pentapeptide (UDP-MurNAc-pentapeptide), a substrate required for synthesis of the peptidoglycans which constitute a key element of the Mtb cell wall. As the Mur enzymes have no counterparts in the eukaryotic cell, they constitute a great target against TB infections.

With the aim of discovering new drugs against the Mur enzymes, the authors developed a one-pot assay to reconstruct the entire Mur pathway in vitro, allowing the screening of multiple molecules against the Mur pathway in a single assay. In this assay the product of the first reaction became the substrate of the following one. To determine the efficiency of each reaction, our PiColorLock™ Phosphate Detection System was used to measure the uncoupled hydrolysis of ATP by Mur enzymes.
Finally, after validating the one-pot assay as a method to recreate the Mur pathway in vitro, the authors identified at least two new potential multi-target drugs against the Mur family. Further studies using the one-pot assay could result in discovering new drugs to be used in multi-targeted therapy. Read more

Key features and advantages of using Innova’s PiColorLock™ Phosphate Detection System:

  • Colorimetric assay, non-radioactive
  • Compatible with almost any assay buffer making it flexible for a variety of applications
  • Unique accelerator (fast reaction) and stabilizer (no precipitation even after several hours)
  • Wide linear range (no inhibition due to high protein concentration)
  • Long shelf-life due to stable reagent formulation

Find out more about our range of products for measuring inorganic phosphate (Pi)