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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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Article information


Submitted
31 Oct 2017
Accepted
12 Jan 2018
First published
15 Jan 2018

This article is Open Access

Anal. Methods, 2018,10, 646-651
Article type
Paper

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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    [Original citation] - Published by The Royal Society of Chemistry.

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