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

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http://hdl.handle.net/10138/237234

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Vidilaseris , K , Kellosalo , J & Goldman , A 2018 , ' A high-throughput method for orthophosphate determination of thermostable membrane-bound pyrophosphatase activity ' , Analytical Methods , vol. 10 , no. 6 , pp. 646-651 . https://doi.org/10.1039/c7ay02558k

Titel: A high-throughput method for orthophosphate determination of thermostable membrane-bound pyrophosphatase activity
Författare: Vidilaseris, Keni; Kellosalo, Juho; Goldman, Adrian
Upphovmannens organisation: Biosciences
Institute of Biotechnology
Biochemistry and Biotechnology
Datum: 2018-02-14
Språk: eng
Sidantal: 6
Tillhör serie: Analytical Methods
ISSN: 1759-9660
DOI: https://doi.org/10.1039/c7ay02558k
Permanenta länken (URI): http://hdl.handle.net/10138/237234
Abstrakt: 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.
Subject: LABILE ORGANIC PHOSPHATE
COLI INORGANIC PYROPHOSPHATASE
COLORIMETRIC DETERMINATION
THERMOTOGA-MARITIMA
HETEROLOGOUS EXPRESSION
PUMPING PYROPHOSPHATASE
CRYSTAL-STRUCTURE
ASSAY
ACIDOCALCISOMES
PURIFICATION
116 Chemical sciences
Referentgranskad: Ja
Licens: cc_by_nc
Användningsbegränsning: openAccess


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