Evidence map›Paper›PMID 35525556›Full record

ArticleMethods in enzymology2022

Efficient expression, purification, and visualization by cryo-EM of unliganded near full-length HER3.

Devan Diwanji, Raphael Trenker, Natalia Jura, Kliment A Verba

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

Article in Methods in enzymology, 2022. 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
1.5field-weighted citation impact, top 19% 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, 1 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

4 authors at 2 institutions in 1 country.

Devan DiwanjiCardiovascular Research Institute, University of California San Francisco, San Francisco, CA, United States; Medical Scientist Training Program, University of California San Francisco, San Francisco, CA, United States.
Raphael TrenkerCardiovascular Research Institute, University of California San Francisco, San Francisco, CA, United States.
Natalia JuraCardiovascular Research Institute, University of California San Francisco, San Francisco, CA, United States; Quantitative Biosciences Institute (QBI), University of California San Francisco, San Francisco, CA, United States; Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, CA, United States. Electronic address: natalia.jura@ucsf.edu.
Kliment A VerbaQuantitative Biosciences Institute (QBI), University of California San Francisco, San Francisco, CA, United States; Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, CA, United States. Electronic address: kliment.verba@ucsf.edu.
University of California, San Francisco · USQuantitative BioSciences · US

Funding

Using Networks to Seed Hierarchical Whole-cell Models of CancerU54CA209891 · NCI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI KROGAN, NEVAN J · 2017 to 2021
$10.9M
Structural and Functional Studies of HER ReceptorsR35GM139636 · NIGMS · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI Natalia Jura · 2021 to 2026
$2.5M
Investigating oncogenic mutations and regulatory mechanisms of HER2 by electron microscopyF30CA247147 · NCI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI DIWANJI, DEVAN · 2020 to 2023
$163k
NCI NIH HHS F30 CA247147NCI NIH HHS U54 CA209891NIGMS NIH HHS R35 GM139636
6 · The paper itself

Abstract

Biochemical analyses of membrane receptor kinases have been limited by challenges in obtaining sufficient homogeneous receptor samples for downstream structural and biophysical characterization. Here, we report a suite of methods for the efficient expression, purification, and visualization by cryo-electron microscopy (cryo-EM) of near full-length Human Epidermal Growth Factor Receptor 3 (HER3), a receptor tyrosine pseudokinase, in the unliganded state. Through transient mammalian cell expression, a two-step purification with detergent exchange into lauryl maltose neopentyl glycol (LMNG), and freezing devoid of background detergent micelle, we obtained ~6Å reconstructions of the ~60kDa fully-glycosylated unliganded extracellular domain of HER3 from just 30mL of suspension culture. The reconstructions reveal previously unappreciated extracellular domain dynamics and glycosylation sites.

Indexed as

DetergentsMicellesAnimalsCryoelectron MicroscopyHumansMammalsDetergentsMicellesCryo-electron microscopyHER receptorsHuman epidermal growth factor receptorsMembrane proteinsReceptor tyrosine kinases

Identifiers

PMID35525556
PMCPMC9288109
OpenAlexW4225502804

What Socratic holds

Textmetadata
LicenceTDM
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 Socratic graph.