Evidence map›Paper›PMID 28253433›Full record

ArticleCurrent protocols in chemical biology2017

UbFluor: A Fluorescent Thioester to Monitor HECT E3 Ligase Catalysis.

David T Krist, Peter K Foote, Alexander V Statsyuk

Open access · greenAbstract read
In one paragraph

Article in Current protocols in chemical biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
0.6field-weighted citation impact, top 32% 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

6 citing papers in PubMed, 7 citations in OpenAlex.

  1. Review
  2. Chemical Tools for Probing the Ub/Ubl Conjugation Cascades.Chembiochem : a European journal of chemical biology · 2025
    Review
  3. Targeting TRIM Proteins: A Quest towards Drugging an Emerging Protein Class.Chembiochem : a European journal of chemical biology · 2021
    Review
  4. Review
  5. Article
  6. 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

3 authors at 1 institution in 1 country.

David T KristNorthwestern University, Department of Chemistry, Chemistry of Life Processes Institute, Evanston, Illinois.
Peter K FooteNorthwestern University, Department of Chemistry, Chemistry of Life Processes Institute, Evanston, Illinois.
Alexander V StatsyukNorthwestern University, Department of Chemistry, Chemistry of Life Processes Institute, Evanston, Illinois.
Northwestern University · US

Funding

Molecular Biophysics Training Program at Northwestern UniversityT32GM008382 · NIGMS · NORTHWESTERN UNIVERSITY · PI RADHAKRISHNAN, ISHWAR · 1990 to 2020
$4.3M
Pharmacological Inhibitors of Nedd4 ubiquitin ligase as anticancer therapeuticsR01GM115632 · NIGMS · UNIVERSITY OF HOUSTON · PI STATSYUK, ALEXANDER V · 2015 to 2019
$1.5M
NIGMS NIH HHS R01 GM115632NIGMS NIH HHS T32 GM008382
6 · The paper itself

Abstract

HECT E3 ubiquitin ligases (∼28 are known) are associated with many phenotypes in eukaryotes and are important drug targets. However, assays used to screen for small molecule inhibitors of HECT E3s are complex and require ATP, Ub, E1, E2, and HECT E3 enzymes, producing three covalent thioester enzyme intermediates E1∼Ub, E2∼Ub, and HECT E3∼Ub (where ∼ indicates a thioester bond), and mixtures of polyubiquitin chains. To reduce the complexity of the assay, we developed a novel class of fluorescent probes, UbFluor, that act as mechanistically relevant pseudosubstrates of HECT E3s. These probes undergo a direct transthiolation reaction with the catalytic cysteine of HECT E3s, producing the catalytically active HECT E3∼Ub thioester accompanied by fluorophore release. Thus, a fluorescence polarization assay can continuously monitor UbFluor consumption by HECT E3s, and changes in UbFluor consumption rendered by biochemical point mutations or small molecule modulation of HECT E3 activity. © 2017 by John Wiley & Sons, Inc.

Indexed as

HumansModels, MolecularUbiquitinUbiquitin-Protein LigasesUbiquitinUbiquitin-Protein Ligasesfluorescence polarizationHECT ligasethioesterubiquitin

Identifiers

PMID28253433
PMCPMC5335898
OpenAlexW2781979290

What OpenQuestion 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 OpenQuestion graph.