Evidence map›Paper›PMID 34298920›Full record

ArticleInternational journal of molecular sciences2021

Combined FCS and PCH Analysis to Quantify Protein Dimerization in Living Cells.

Laura M Nederveen-Schippers, Pragya Pathak, Ineke Keizer-Gunnink, Adrie H Westphal, Peter J M van Haastert, Jan Willem Borst, Arjan Kortholt, Victor Skakun

Open access · goldAbstract read
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 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
1.5field-weighted citation impact, top 14% 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

2 citing papers in PubMed, 7 citations in OpenAlex.

  1. Article
  2. 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

8 authors at 3 institutions in 2 countries.

Laura M Nederveen-SchippersDepartment of Cell Biochemistry, University of Groningen, 9747 AG Groningen, The Netherlands.
Pragya PathakDepartment of Cell Biochemistry, University of Groningen, 9747 AG Groningen, The Netherlands.ORCID 0000-0002-7259-6558
Ineke Keizer-GunninkDepartment of Cell Biochemistry, University of Groningen, 9747 AG Groningen, The Netherlands.
Adrie H WestphalLaboratory of Biochemistry, Wageningen University & Research, 6708 WE Wageningen, The Netherlands.
Peter J M van HaastertDepartment of Cell Biochemistry, University of Groningen, 9747 AG Groningen, The Netherlands.
Jan Willem BorstLaboratory of Biochemistry, Wageningen University & Research, 6708 WE Wageningen, The Netherlands.
Arjan KortholtDepartment of Cell Biochemistry, University of Groningen, 9747 AG Groningen, The Netherlands.ORCID 0000-0001-8174-6397
Victor SkakunDepartment of Systems Analysis and Computer Simulation, Belarusian State University, 220030 Minsk, Belarus.ORCID 0000-0003-0880-4188
University of Groningen · NLWageningen University & Research · NLBelarusian State University · BY

Funding

Nederlandse Organisatie voor Wetenschappelijk Onderzoek 723.012.108
6 · The paper itself

Abstract

Protein dimerization plays a crucial role in the regulation of numerous biological processes. However, detecting protein dimers in a cellular environment is still a challenge. Here we present a methodology to measure the extent of dimerization of GFP-tagged proteins in living cells, using a combination of fluorescence correlation spectroscopy (FCS) and photon counting histogram (PCH) analysis of single-color fluorescence fluctuation data. We named this analysis method brightness and diffusion global analysis (BDGA) and adapted it for biological purposes. Using cell lysates containing different ratios of GFP and tandem-dimer GFP (diGFP), we show that the average brightness per particle is proportional to the fraction of dimer present. We further adapted this methodology for its application in living cells, and we were able to distinguish GFP, diGFP, as well as ligand-induced dimerization of FKBP12 (FK506 binding protein 12)-GFP. While other analysis methods have only sporadically been used to study dimerization in living cells and may be prone to errors, this paper provides a robust approach for the investigation of any cytosolic protein using single-color fluorescence fluctuation spectroscopy.

Indexed as

Cells, CulturedCytosolDictyosteliumDiffusionDimerizationFluorescenceGreen Fluorescent ProteinsLigandsPhotonsProtein MultimerizationProteinsSpectrometry, FluorescenceGreen Fluorescent ProteinsLigandsProteinsbrightness and diffusion global analysisDictyostelium discoideumdimeric proteinFK506 binding protein 12fluorescence correlation spectroscopyfluorescence fluctuation spectroscopyGFPphoton counting histogram

Identifiers

PMID34298920
PMCPMC8307594
OpenAlexW3180687337

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.