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Related Concept Videos

Proteomics01:33

Proteomics

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 proteomics...

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Glycopeptide Capture for Cell Surface Proteomics
10:11

Glycopeptide Capture for Cell Surface Proteomics

Published on: May 9, 2014

GOFAST: an integrated approach for efficient and comprehensive membrane proteome analysis.

Yanbao Yu1, Ling Xie, Harsha P Gunawardena

  • 1Department of Biochemistry & Biophysics, University of North Carolina at Chapel Hill, North Carolina 27599, United States.

Analytical Chemistry
|October 4, 2012
PubMed
Summary

We developed GELFrEE Optimized FASP Technology (GOFAST) for comprehensive membrane proteomics. This method enhances membrane protein identification and recovery, yielding the largest leukemia membrane proteome to date.

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Area of Science:

  • Proteomics
  • Cell Biology
  • Biochemistry

Background:

  • Membrane proteomics faces challenges in protein separation and proteolysis resistance, leading to low recovery and identification rates.
  • Existing methods often struggle with comprehensive analysis of membrane proteins.

Purpose of the Study:

  • To introduce a novel integrated approach, GELFrEE Optimized FASP Technology (GOFAST), for large-scale and comprehensive membrane protein analysis.
  • To overcome limitations in membrane protein separation and identification.

Main Methods:

  • Integrated approach combining gel-eluted liquid fraction entrapment electrophoresis (GELFrEE), filter-aided sample preparation (FASP), and microwave-assisted on-filter enzymatic digestion.
  • Analysis of the membrane fraction from K562 leukemia cell line.

Main Results:

  • Identified 2,090 proteins from the K562 membrane fraction.
  • 37% of identified proteins are annotated as membrane proteins, representing the largest membrane proteome of leukemia cells reported.
  • Demonstrated high protein recovery and maximized MS-detectable sequence coverage.

Conclusions:

  • GOFAST offers a powerful and comprehensive approach for large-scale membrane proteomics.
  • The method enhances efficiency by combining GELFrEE, FASP, and microwave-assisted digestion, minimizing sample loss.
  • GOFAST shows potential for identifying alternative splicing products in membrane proteomes.