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Related Concept Videos

Proteomics01:33

Proteomics

9.9K
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
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Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
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Digging More Missing Proteins Using an Enrichment Approach with ProteoMiner.

Siqi Li1, Yanbin He1, Zhilong Lin1

  • 1BGI-Shenzhen , Beishan Industrial Zone 11th building, Yantian District, Shenzhen, Guangdong 518083, China.

Journal of Proteome Research
|September 30, 2017
PubMed
Summary

The Human Proteome Project aims to map all human proteins. This study demonstrates that ProteoMiner enrichment effectively identifies previously undetected low-abundance missing proteins in human tissues.

Keywords:
LC−MS/MSProteoMiner protein enrichment kitTriton X-100human tissuemissing proteins

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Area of Science:

  • Proteomics
  • Human Biology
  • Biochemistry

Background:

  • The Human Proteome Project (HPP) endeavors to map all human proteins to advance biological understanding and medical applications.
  • Despite advanced techniques, 2563 missing proteins (MPs) remain undetected, hindering comprehensive proteome coverage.

Purpose of the Study:

  • To investigate the efficacy of low-abundance protein enrichment for discovering missing proteins.
  • To evaluate the ProteoMiner technique for identifying previously undetected human proteins.

Main Methods:

  • Utilized ProteoMiner, an equalizing technique, to enrich low-abundance proteins by reducing high-abundance ones.
  • Employed triton X-100/TBS buffer extraction, ProteoMiner enrichment, and peptide fractionation for protein analysis.
  • Validated identified missing proteins using Parallel Reaction Monitoring (PRM) assays against synthetic peptides.

Main Results:

  • Identified 20 missing proteins (MPs), including membrane/secreted and nucleus proteins, from four human tissues.
  • Confirmed 15 of these MPs using two unique peptides (≥9 amino acids) via PRM assay.
  • Successfully identified at least two non-nested unique peptides (≥8 amino acids) for each reported MP.

Conclusions:

  • ProteoMiner enrichment is a powerful strategy for the discovery of low-abundance missing proteins.
  • This approach significantly enhances the detection of previously unmapped human proteins.
  • The findings contribute to a more complete human proteome map and potential diagnostic/therapeutic targets.