Evidence map›Paper›PMID 34768989›Full record

ArticleInternational journal of molecular sciences2021

Semi-Automated High-Throughput Substrate Screening Assay for Nucleoside Kinases.

Katja F Hellendahl, Maryke Fehlau, Sebastian Hans, Peter Neubauer, Anke Kurreck

Open access · goldAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in International journal of molecular sciences, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
0.5field-weighted citation impact, top 37% 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

1 citing paper in PubMed, 6 citations in OpenAlex.

  1. Article
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

5 authors at 1 institution in 1 country.

Katja F HellendahlChair of Bioprocess Engineering, Faculty III Process Sciences, Institute of Biotechnology, Technische Universität Berlin, Ackerstraße 76, 13355 Berlin, Germany.
Maryke FehlauChair of Bioprocess Engineering, Faculty III Process Sciences, Institute of Biotechnology, Technische Universität Berlin, Ackerstraße 76, 13355 Berlin, Germany.
Sebastian HansChair of Bioprocess Engineering, Faculty III Process Sciences, Institute of Biotechnology, Technische Universität Berlin, Ackerstraße 76, 13355 Berlin, Germany.ORCID 0000-0002-5346-869X
Peter NeubauerChair of Bioprocess Engineering, Faculty III Process Sciences, Institute of Biotechnology, Technische Universität Berlin, Ackerstraße 76, 13355 Berlin, Germany.ORCID 0000-0002-1214-9713
Anke KurreckChair of Bioprocess Engineering, Faculty III Process Sciences, Institute of Biotechnology, Technische Universität Berlin, Ackerstraße 76, 13355 Berlin, Germany.
Technische Universität Berlin · DE

Funding

Deutsche Forschungsgemeinschaft 392246628
6 · The paper itself

Abstract

Nucleoside kinases (NKs) are key enzymes involved in the in vivo phosphorylation of nucleoside analogues used as drugs to treat cancer or viral infections. Having different specificities, the characterization of NKs is essential for drug design and nucleotide analogue production in an in vitro enzymatic process. Therefore, a fast and reliable substrate screening method for NKs is of great importance. Here, we report on the validation of a well-known luciferase-based assay for the detection of NK activity in a 96-well plate format. The assay was semi-automated using a liquid handling robot. Good linearity was demonstrated (r² > 0.98) in the range of 0-500 µM ATP, and it was shown that alternative phosphate donors like dATP or CTP were also accepted by the luciferase. The developed high-throughput assay revealed comparable results to HPLC analysis. The assay was exemplarily used for the comparison of the substrate spectra of four NKs using 20 (8 natural, 12 modified) substrates. The screening results correlated well with literature data, and additionally, previously unknown substrates were identified for three of the NKs studied. Our results demonstrate that the developed semi-automated high-throughput assay is suitable to identify best performing NKs for a wide range of substrates.

Indexed as

Adenosine TriphosphateAnimalsDrosophila melanogasterDrug Evaluation, PreclinicalHigh-Throughput Screening AssaysHumansLuciferasesNucleosidesPhosphorylationPhosphotransferasesSubstrate SpecificityAdenosine TriphosphateLuciferasesnucleoside phosphotransferaseNucleosidesPhosphotransferasesbiocatalysishigh throughputHPLCluciferase assayluminescent assaynucleoside analoguesnucleoside kinasenucleotiderobotscreening

Identifiers

PMID34768989
PMCPMC8584170
OpenAlexW3209396329

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

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.