Evidence map›Paper›PMID 37026813›Full record

ArticleCurrent protocols2023

Proteasomes: Isolation and Activity Assays.

Yanjie Li, Robert J Tomko, Mark Hochstrasser

Open access · greenAbstract read
In one paragraph

Article in Current protocols, 2023. 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
0.2field-weighted citation impact, top 43% 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, 3 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

3 authors at 2 institutions in 1 country.

Yanjie LiDepartment of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut.
Robert J TomkoDepartment of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, Florida.
Mark HochstrasserDepartment of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut.
Yale University · USFlorida State University · US

Funding

Mechanisms of Cell Regulation and Manipulation by the Ubiquitin SystemR35GM136325 · NIGMS · YALE UNIVERSITY · PI Mark W Hochstrasser · 2020 to 2026
$6.5M
Engagement and Communication Between Proteasonal SubcomplexesR01GM118600 · NIGMS · FLORIDA STATE UNIVERSITY · PI ROBERT JOSEPH TOMKO · 2017 to 2026
$2.7M
Investigation of the proteasome assembly landscapeR01GM144550 · NIGMS · FLORIDA STATE UNIVERSITY · PI SUO, ZUCAI, TOMKO, ROBERT JOSEPH · 2022 to 2025
$1.7M
NIGMS NIH HHS R01 GM118600NIGMS NIH HHS R01 GM144550NIGMS NIH HHS R35 GM136325NIH HHS R01 GM118600NIH HHS R01 GM144550NIH HHS R35 GM136325
6 · The paper itself

Abstract

In eukaryotes, damaged or unneeded proteins are typically degraded by the ubiquitin-proteasome system. In this system, the protein substrate is often first covalently modified with a chain of ubiquitin polypeptides. This chain serves as a signal for delivery to the 26S proteasome, a 2.5-MDa, ATP-dependent multisubunit protease complex. The proteasome consists of a barrel-shaped 20S core particle (CP) that is capped on one or both of its ends by a 19S regulatory particle (RP). The RP is responsible for recognizing the substrate, unfolding it, and translocating it into the CP for destruction. Here we describe simple, one-step purification schemes for isolating the 26S proteasome and its 19S RP and 20S CP subcomplexes from the yeast Saccharomyces cerevisiae. A gel filtration step can be added to further enhance purity. We also describe assays for measuring ubiquitin-dependent and ubiquitin-independent proteolytic activity in vitro. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Purification of active 26S proteasomes Support Protocol 1: Growth of yeast strains and production of yeast cell powder Support Protocol 2: Regeneration of anti-flag M2 affinity gel Basic Protocol 2: Purification of the 19S regulatory particle (RP) Basic Protocol 3: Purification of active 20S CP Basic Protocol 4: In-gel peptidase activity assay for 20S CP and 26S proteasomes Basic Protocol 5: In-solution peptidase activity assay for 20S and 26S proteasomes Basic Protocol 6: Measuring degradation of polyubiquitinated SIC1

Indexed as

Proteasome Endopeptidase ComplexYeast, DriedCytoplasmSaccharomyces cerevisiaeUbiquitinProteasome Endopeptidase ComplexUbiquitinATPaseproteasomeproteolytic activitypurificationubiquitin

Identifiers

PMID37026813
PMCPMC10337785
OpenAlexW4362692566

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.