Evidence map›Paper›PMID 29388937›Full record

ArticleNature protocols2018

Characterization of homodimer interfaces with cross-linking mass spectrometry and isotopically labeled proteins.

Diogo B Lima, John T Melchior, Jamie Morris, Valmir C Barbosa, Julia Chamot-Rooke, Mariana Fioramonte, Tatiana A C B Souza, Juliana S G Fischer, Fabio C Gozzo, Paulo C Carvalho and 1 more

Abstract read
PubMed Publisher
In one paragraph

Article in Nature protocols, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.

0numbers the graph read from it
0cells of the map it votes in
33citing papers in PubMed
–field-weighted citation impact
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

33 citing papers in PubMed.

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  20. Biological Applications for LC-MS-Based Proteomics.Advances in experimental medicine and biology · 2021
    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

11 authors.

Diogo B LimaMass Spectrometry for Biology Unit, CNRS USR 2000, Institut Pasteur, Paris, France.
John T MelchiorPathology and Laboratory Medicine, University of Cincinnati, Cincinnati, Ohio, USA.
Jamie MorrisPathology and Laboratory Medicine, University of Cincinnati, Cincinnati, Ohio, USA.
Valmir C BarbosaSystems Engineering and Computer Science Program, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.ORCID 0000-0003-1010-7170
Julia Chamot-RookeMass Spectrometry for Biology Unit, CNRS USR 2000, Institut Pasteur, Paris, France.
Mariana FioramonteDalton Mass Spectrometry Laboratory, University of Campinas, São Paulo, Brazil.
Tatiana A C B SouzaGroup for Computational Mass Spectrometry & Proteomics, Carlos Chagas Institute, Fiocruz Paraná, Brazil.
Juliana S G FischerGroup for Computational Mass Spectrometry & Proteomics, Carlos Chagas Institute, Fiocruz Paraná, Brazil.
Fabio C GozzoDalton Mass Spectrometry Laboratory, University of Campinas, São Paulo, Brazil.
Paulo C CarvalhoGroup for Computational Mass Spectrometry & Proteomics, Carlos Chagas Institute, Fiocruz Paraná, Brazil.ORCID 0000-0001-6530-3350
W Sean DavidsonPathology and Laboratory Medicine, University of Cincinnati, Cincinnati, Ohio, USA.

Funding

A high-resolution structural approach to understanding HDL biogenesisR01GM098458 · NIGMS · UNIVERSITY OF CINCINNATI · PI DAVIDSON, W SEAN · 2011 to 2014
$1.2M
NIH HHS R01 GM098458
6 · The paper itself

Abstract

Cross-linking coupled with mass spectrometry (XL-MS) has emerged as a powerful strategy for the identification of protein-protein interactions, characterization of interaction regions, and obtainment of structural information on proteins and protein complexes. In XL-MS, proteins or complexes are covalently stabilized with cross-linkers and digested, followed by identification of the cross-linked peptides by tandem mass spectrometry (MS/MS). This provides spatial constraints that enable modeling of protein (complex) structures and regions of interaction. However, most XL-MS approaches are not capable of differentiating intramolecular from intermolecular links in multimeric complexes, and therefore they cannot be used to study homodimer interfaces. We have recently developed an approach that overcomes this limitation by stable isotope-labeling of one of the two monomers, thereby creating a homodimer with one 'light' and one 'heavy' monomer. Here, we describe a step-by-step protocol for stable isotope-labeling, followed by controlled denaturation and refolding in the presence of the wild-type protein. The resulting light-heavy dimers are cross-linked, digested, and analyzed by mass spectrometry. We show how to quantitatively analyze the corresponding data with SIM-XL, an XL-MS software with a module tailored toward the MS/MS data from homodimers. In addition, we provide a video tutorial of the data analysis with this protocol. This protocol can be performed in ∼14 d, and requires basic biochemical and mass spectrometry skills.

Indexed as

Amino Acid SequenceCross-Linking ReagentsIsotope LabelingPeptidesProtein ConformationProteinsSoftwareTandem Mass SpectrometryCross-Linking ReagentsPeptidesProteins

Identifiers

PMID29388937

What OpenQuestion holds

Textmetadata
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.