Evidence map›Paper›PMID 31528665›Full record

ArticleBio-protocol2019

Cross-linking, Immunoprecipitation and Proteomic Analysis to Identify Interacting Proteins in Cultured Cells.

Hao Wang, Meiling He, Belinda Willard, Qingyu Wu

Open access · hybridAbstract read
In one paragraph

Article in Bio-protocol, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
0.9field-weighted citation impact, top 30% 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

8 citing papers in PubMed, 13 citations in OpenAlex.

  1. Article
  2. Article
  3. Article
  4. Article
  5. Article
  6. In Vivo Protein Cross-Linking and Coimmunoprecipitation in Haloferax volcanii.Methods in molecular biology (Clifton, N.J.) · 2022
    Article
  7. Article
  8. 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 2 countries.

Hao WangDepartment of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China.
Meiling HeCyrus Tang Hematology Center, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Prevention, Soochow University, Suzhou, China.
Belinda WillardProteomics Core, Lerner Research Institute, Cleveland Clinic, Cleveland, USA.
Qingyu WuDepartment of Cardiovascular & Metabolic Sciences, Lerner Research Institute, Cleveland Clinic, Cleveland, USA.
Cleveland Clinic Lerner College of Medicine · USSoochow University · CN

Funding

Molecular Mechanisms In Corin ActivationR01HL126697 · NHLBI · CLEVELAND CLINIC LERNER COM-CWRU · PI WU, QINGYU · 2015 to 2018
$1.6M
LTQ-Orbitrap Velos instrumentS10RR031537 · NCRR · CLEVELAND CLINIC LERNER COM-CWRU · PI WILLARD, BELINDA BELLE · 2011 to 2011
$600k
Trophoblasts in Preeclampsia Associated Thrombotic ComplicationsR01HD093727 · NICHD · CLEVELAND CLINIC LERNER COM-CWRU · PI WU, QINGYU · 2018 to 2019
$559k
NCRR NIH HHS S10 RR031537NHLBI NIH HHS R01 HL126697NICHD NIH HHS R01 HD093727
6 · The paper itself

Abstract

Extracellular expression is essential for the function of secreted and cell surface proteins. Proper intracellular trafficking depends on protein interactions in multiple subcellular compartments. Co-immunoprecipitation and the yeast two-hybrid system are commonly used to investigate protein-protein interactions. These methods, however, depend on high-affinity protein interactions. In many glycoproteins, glycans are important for protein intracellular trafficking and extracellular expression. If glycoprotein interactions are transient and relatively weak, it may be challenging to use co-immunoprecipitation or the two-hybrid system to identify glycoprotein-binding partners. To circumvent this problem, protein cross-linking can be applied first to immobilize the transient and/or low-affinity protein interactions. Here we describe a protocol of protein cross-linking, co-immunoprecipitation, and proteomic analysis, which was used to identify endoplasmic reticulum (ER) chaperones critical for the folding and ER exiting of N-glycosylated serine proteases in human embryonic kidney (HEK) 293 cells. This approach can be used to identify other protein interactions in a variety of cells.

Indexed as

CorinImmunoprecipitationN-glycosylationProtein cross-linkingProtein-protein interactionProteomic analysisSerine protease

Identifiers

PMID31528665
PMCPMC6746336
OpenAlexW2949051640

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.