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Issue 6, 2018
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A high-throughput method for orthophosphate determination of thermostable membrane-bound pyrophosphatase activity

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Abstract

Membrane-bound pyrophosphatases (mPPases) are homodimeric integral membrane proteins that hydrolyse pyrophosphate into orthophosphates coupled to the active transport of protons or sodium ions across membranes. They occur in bacteria, archaea, plants, and protist parasites. As they are essential in protist parasites and there are no homologous proteins in animals and humans, these enzymes represent an excellent drug target for treating protistal diseases. Experimental screening to find drug candidates is an important step to discover new hit compounds. For that, a cheap, simple, and robust assay is needed. Here we report the application of the molybdenum blue reaction method for a medium throughput microplate activity assay of the hyperthermophilic bacterium Thermotoga maritima mPPase and the possible application of the assay to screen inhibitors of membrane-bound pyrophosphatases.

Graphical abstract: A high-throughput method for orthophosphate determination of thermostable membrane-bound pyrophosphatase activity

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Publication details

The article was received on 31 Oct 2017, accepted on 12 Jan 2018 and first published on 15 Jan 2018


Article type: Paper
DOI: 10.1039/C7AY02558K
Anal. Methods, 2018,10, 646-651
  • Open access: Creative Commons BY-NC license
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    A high-throughput method for orthophosphate determination of thermostable membrane-bound pyrophosphatase activity

    K. Vidilaseris, J. Kellosalo and A. Goldman, Anal. Methods, 2018, 10, 646
    DOI: 10.1039/C7AY02558K

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