Cell-free expression, purification, and ligand-binding analysis of Drosophila melanogaster olfactory receptors DmOR67a, DmOR85b and DmORCO.

Lotta Tollstoy Tegler, Karolina Corin, Julia Hillger, Brooke Wassie, Yanmei Yu, Shuguang Zhang

Open access · goldAbstract read

In one paragraph

Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.

0numbers the graph read from it

0cells of the map it votes in

7citing papers in PubMed

1.8field-weighted citation impact, top 17% 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

7 citing papers in PubMed, 21 citations in OpenAlex.

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TRPA1-FGFR2 binding event is a regulatory oncogenic driver modulated by miRNA-142-3p.Nature communications · 2017
Article
From Gene to Function: Cell-Free Electrophysiological and Optical Analysis of Ion Pumps in Nanodiscs.Biophysical journal · 2017
Article
Cell-free methods to produce structurally intact mammalian membrane proteins.Scientific reports · 2016
Article
Expression Divergence of Chemosensory Genes between Drosophila sechellia and Its Sibling Species and Its Implications for Host Shift.Genome biology and evolution · 2015
Article
Cell-free expression of a functional pore-only sodium channel.Protein expression and purification · 2015
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

6 authors at 2 institutions in 1 country.

Lotta Tollstoy Tegler1] Center for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts, Cambridge, MA 02139-4307, USA [2] Division of Chemistry, Department of Physics, Chemistry and Biology, Linkoping University, SE-581 83 Linkoping, Sweden.

Karolina Corin1] Center for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts, Cambridge, MA 02139-4307, USA [2] Biomedical Engineering Research Group, School of Electrical and Information Engineering University of the Witwatersrand, Johannesburg, South Africa [3] Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa.

Julia HillgerCenter for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts, Cambridge, MA 02139-4307, USA.

Brooke WassieCenter for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts, Cambridge, MA 02139-4307, USA.

Yanmei Yu1] Center for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts, Cambridge, MA 02139-4307, USA [2] College of Electronics and Information Engineering, Sichuan University, Chengdu, China 610065.

Shuguang ZhangCenter for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts, Cambridge, MA 02139-4307, USA.

MIT-Harvard Center for Ultracold Atoms · USMassachusetts Institute of Technology · US

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Insects transmit numerous devastating diseases, including malaria, dengue fever, and sleeping sickness. Olfactory cues guide insects to their hosts, and are thus responsible for disease transmission. Understanding the molecular basis of insect olfaction could facilitate the development of interventions. The first step is to heterologously overexpress and purify insect olfactory receptors (ORs). This is challenging, as ORs are membrane proteins. Here, we show that insect ORs and their co-receptor can be expressed in an E. coli cell-free system. After immunoaffinity chromatography, the ORs are ~95% pure, and up to 1 mg/10 ml reaction is obtained. Circular dichroism together with microscale thermophoresis indicate that each receptor is properly folded, and can bind its respective ligand. This is the first time insect ORs have been expressed in an E. coli system. The methods described here could facilitate future structure-function studies, which may aid in developments to alleviate the suffering of millions caused by insect-transmitted diseases.

Indexed as

AnimalsCell-Free SystemDrosophila melanogasterDrosophila ProteinsEscherichia coliGene ExpressionLigandsOlfactory Receptor NeuronsProtein BindingReceptors, Cell SurfaceReceptors, OdorantDrosophila ProteinsLigandsOr67d protein, DrosophilaOrco protein, DrosophilaReceptors, Cell SurfaceReceptors, Odorant

Identifiers

PMID25597985

DOI10.1038/srep07867

PMCPMC4297953

OpenAlexW2025641578

What OpenQuestion holds

Textmetadata

LicenceCC BY-NC-SA

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.