Evidence map›Paper›PMID 26482925›Full record

ArticlePurinergic signalling2015

Mass spectrometry-based ligand binding assays on adenosine A1 and A2A receptors.

A Massink, M Holzheimer, A Hölscher, J Louvel, D Guo, G Spijksma, T Hankemeier, A P IJzerman

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PubMed Full text at PMC Publisher
In one paragraph

Article in Purinergic signalling, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
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

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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

4 citing papers in PubMed.

  1. Review
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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

8 authors.

A MassinkDivision of Medicinal Chemistry, LACDR, Leiden University, Leiden, The Netherlands.
M HolzheimerDivision of Medicinal Chemistry, LACDR, Leiden University, Leiden, The Netherlands.
A HölscherDivision of Medicinal Chemistry, LACDR, Leiden University, Leiden, The Netherlands.
J LouvelDivision of Medicinal Chemistry, LACDR, Leiden University, Leiden, The Netherlands.
D GuoDivision of Medicinal Chemistry, LACDR, Leiden University, Leiden, The Netherlands.
G SpijksmaDivision of Analytical Biosciences, LACDR, Leiden University, Leiden, The Netherlands.
T HankemeierDivision of Analytical Biosciences, LACDR, Leiden University, Leiden, The Netherlands.
A P IJzermanDivision of Medicinal Chemistry, LACDR, Leiden University, Leiden, The Netherlands. ijzerman@lacdr.leidenuniv.nl.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Conventional methods to measure ligand-receptor binding parameters typically require radiolabeled ligands as probes. Despite the robustness of radioligand binding assays, they carry inherent disadvantages in terms of safety precautions, expensive synthesis, special lab requirements, and waste disposal. Mass spectrometry (MS) is a method that can selectively detect ligands without the need of a label. The sensitivity of MS equipment increases progressively, and currently, it is possible to detect low ligand quantities that are usually found in ligand binding assays. We developed a label-free MS ligand binding (MS binding) assay on the adenosine A(1) and A(2A) receptors (A(1)AR and A(2A)AR), which are well-characterized members of the class A G protein-coupled receptor (GPCR) family. Radioligand binding assays for both receptors are well established, and ample data is available to compare and evaluate the performance of an MS binding assay. 1,3-Dipropyl-8-cyclopentyl-xanthine (DPCPX) and 4-(2-((7-amino-2-(furan-2-yl)-[1,2,4]triazolo[1,5-a]-[1,3,5]triazin-5-yl)amino)ethyl)phenol (ZM-241,385) are high-affinity ligands selective for the A(1)AR and A(2A)AR, respectively. To proof the feasibility of MS binding on the A(1)AR and A(2A)AR, we first developed an MS detection method for unlabeled DPCPX and ZM-241,385. To serve as internal standards, both compounds were also deuterium-labeled. Subsequently, we investigated whether the two unlabeled compounds could substitute for their radiolabeled counterparts as marker ligands in binding experiments, including saturation, displacement, dissociation, and competition association assays. Furthermore, we investigated the accuracy of these assays if the use of internal standards was excluded. The results demonstrate the feasibility of the MS binding assay, even in the absence of a deuterium-labeled internal standard, and provide great promise for the further development of label-free assays based on MS for other GPCRs.

Indexed as

AnimalsCell MembraneCHO CellsCricetinaeCricetulusDeuteriumHumansIsotope LabelingLigandsMass SpectrometryRadioligand AssayReceptor, Adenosine A1Receptor, Adenosine A2AReceptors, G-Protein-CoupledReference StandardsReproducibility of Results1,3-dipropyl-8-cyclopentylxanthineDeuteriumLigandsReceptor, Adenosine A1Receptor, Adenosine A2AReceptors, G-Protein-CoupledTriazinesTriazolesXanthinesZM 241385Adenosine receptorDeuterationMass spectrometryMS bindingRadioligand binding

Identifiers

PMID26482925
PMCPMC4648803

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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.