Related Experiment Video

Updated: Jun 1, 2026

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
08:07

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions

Published on: August 2, 2015

Covalent Organic Framework with Acid Precipitation Enhances Microprotein Enrichment and Discovery of

Fengming Li1, Jingchen Sui1, Xiaoqian Wang1

  • 1Department of Chemistry, College of Science, Southern University of Science and Technology, Shenzhen 518055, China.

Analytical Chemistry
|March 3, 2026
PubMed

Insights

We developed novel covalent organic framework (COF)-coated magnetic nanoparticles to efficiently enrich microproteins, which are small proteins often missed by standard methods. This advancement significantly improves microprotein detection and identification in biological samples.

Area of Science:

  • Biochemistry
  • Proteomics
  • Materials Science

Background:

  • Microproteins, encoded by small open reading frames (sORFs), are short polypeptides (<100 amino acids) with unique properties.
  • Detecting microproteins via mass spectrometry is challenging due to their short length and low abundance.
  • Covalent organic frameworks (COFs) show promise for enriching short bioactive peptides.

Purpose of the Study:

  • To develop and validate COF-coated magnetic nanoparticles for enhanced microprotein enrichment from cell lysates.
  • To improve the detection and identification efficiency of microproteins using mass spectrometry.
  • To investigate the role of microproteins in biological processes like ferroptosis.

Main Methods:

  • Synthesis of COF-coated magnetic nanoparticles with a specific pore size (2.72 nm).
  • Utilizing these nanoparticles for microprotein enrichment from cell lysate.
  • Employing mass spectrometry (MS) for microprotein identification and quantification.
  • Combining COF enrichment with acid precipitation for comprehensive microprotein recovery.
  • Applying quantitative proteomics to identify differentially expressed microproteins during ferroptosis.

Main Results:

  • COF-coated nanoparticles identified approximately 5 times more microproteins than uncoated particles.
  • An average of 109 microproteins were identified per MS run (45 min analysis time), with 142 unique microproteins identified across three replicates (FDR < 0.01%).
  • Combining COFs with acid precipitation identified 195 unique microproteins.
  • Seven microproteins were found to be differentially upregulated during ferroptosis, including three novel microproteins confirmed by MS/MS spectra.

Conclusions:

  • COF-coated magnetic nanoparticles provide a robust and efficient tool for microprotein identification.
  • This method significantly enhances the number and reproducibility of microprotein discoveries.
  • The approach facilitates the identification of novel microproteins and their potential roles in biological pathways such as ferroptosis.

More Related Videos

Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides10:27

Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides

Published on: July 14, 2015

Mapping Dysfunctional Protein-Protein Interactions in Disease09:39

Mapping Dysfunctional Protein-Protein Interactions in Disease

Published on: October 24, 2025

Related Concept Videos

Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Precipitation and Co-precipitation01:17

Precipitation and Co-precipitation

Precipitation and coprecipitation methods can be used to separate a mixture of ions in a solution. In qualitative inorganic analysis, ions that form sparingly soluble precipitates with the same reagent are separated based on the differences in solubility products. For example, consider the separation of Cu(II) and Fe(II) ions by precipitation as insoluble sulfides. First, copper(II) sulfide is precipitated by the addition of acidic H2S, where the dissociation of H2S is suppressed. Adding H2S...
Gravimetry: Inorganic And Organic Precipitating Agents00:49

Gravimetry: Inorganic And Organic Precipitating Agents

In gravimetry, the precipitant is chosen carefully to obtain a pure solid that can be easily filtered. Common inorganic precipitants can be used to determine several cations and anions. In some cases, the formation of the same precipitate can be used to determine the cation and the anion. For example, the reaction of barium and chromate ions to give barium chromate is used to determine both barium and chromate. However, precipitates such as hydroxides, oxalates, and metal ammonium phosphates...