Evidence map›Paper›PMID 25785438›Full record

ArticlePloS one2015

Direct determination of phosphatase activity from physiological substrates in cells.

Zhongyuan Ren, Le Duy Do, Géraldine Bechkoff, Saida Mebarek, Nermin Keloglu, Saandia Ahamada, Saurabh Meena, David Magne, Slawomir Pikula, Yuqing Wu and 1 more

Open access · goldAbstract read
In one paragraph

Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
1.6field-weighted citation impact, top 16% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

6 citing papers in PubMed, 18 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

11 authors at 6 institutions in 3 countries.

Zhongyuan RenUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France; State Key Laboratory of Supramolecular Structure and Materials, Jilin University Changchun, 130012, China.
Le Duy DoUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France; Department of Biochemistry, Nencki Institute of Experimental Biology and Polish Academy of Sciences, 02-093 Warsaw, Poland.
Géraldine BechkoffUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
Saida MebarekUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
Nermin KelogluUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
Saandia AhamadaUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
Saurabh MeenaUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
David MagneUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
Slawomir PikulaDepartment of Biochemistry, Nencki Institute of Experimental Biology and Polish Academy of Sciences, 02-093 Warsaw, Poland.
Yuqing WuState Key Laboratory of Supramolecular Structure and Materials, Jilin University Changchun, 130012, China.
René BuchetUniversité de Lyon, Villeurbanne, France; Université Lyon 1, Villeurbanne, France; INSA-Lyon, Villeurbanne, France; CPE Lyon, Villeurbanne, France; Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Villeurbanne, France; CNRS UMR 5246, Villeurbanne, France.
Centre National de la Recherche Scientifique · FRInstitut de Chimie et Biochimie Moléculaires et Supramoléculaires · FRInstitut National des Sciences Appliquées de Lyon · FRÉcole d'Ingénieurs en Chimie et Sciences du Numérique · FRJilin University · CNPolish Academy of Sciences · PL

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

A direct and continuous approach to determine simultaneously protein and phosphate concentrations in cells and kinetics of phosphate release from physiological substrates by cells without any labeling has been developed. Among the enzymes having a phosphatase activity, tissue non-specific alkaline phosphatase (TNAP) performs indispensable, multiple functions in humans. It is expressed in numerous tissues with high levels detected in bones, liver and neurons. It is absolutely required for bone mineralization and also necessary for neurotransmitter synthesis. We provided the proof of concept that infrared spectroscopy is a reliable assay to determine a phosphatase activity in the osteoblasts. For the first time, an overall specific phosphatase activity in cells was determined in a single step by measuring simultaneously protein and substrate concentrations. We found specific activities in osteoblast like cells amounting to 116 ± 13 nmol min(-1) mg(-1) for PPi, to 56 ± 11 nmol min(-1) mg(-1) for AMP, to 79 ± 23 nmol min(-1) mg(-1) for beta-glycerophosphate and to 73 ± 15 nmol min(-1) mg(-1) for 1-alpha-D glucose phosphate. The assay was also effective to monitor phosphatase activity in primary osteoblasts and in matrix vesicles. The use of levamisole--a TNAP inhibitor--served to demonstrate that a part of the phosphatase activity originated from this enzyme. An IC50 value of 1.16 ± 0.03 mM was obtained for the inhibition of phosphatase activity of levamisole in osteoblast like cells. The infrared assay could be extended to determine any type of phosphatase activity in other cells. It may serve as a metabolomic tool to monitor an overall phosphatase activity including acid phosphatases or other related enzymes.

Indexed as

AnimalsCalcification, PhysiologicCell Line, TumorChick EmbryoChondrocytesEnzyme AssaysEnzyme InhibitorsHumansMiceOsteoblastsPhosphoric Monoester HydrolasesSpectrophotometry, InfraredEnzyme InhibitorsPhosphoric Monoester Hydrolases

Identifiers

PMID25785438
PMCPMC4364917
OpenAlexW2086239583

What OpenQuestion holds

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LicenceCC BY
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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.