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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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Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors
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Quantitative proteome profiling of normal human circulating microparticles.

Ole Østergaard1, Christoffer T Nielsen, Line V Iversen

  • 1Department of Clinical Biochemistry and Immunology, Statens Serum Institut, Copenhagen, Denmark.

Journal of Proteome Research
|February 15, 2012
PubMed
Summary

This study quantitatively profiles the normal microparticle (MP) proteome using advanced mass spectrometry. It establishes a baseline MP proteome and a reproducible method for future health and disease research.

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Area of Science:

  • Biochemistry
  • Proteomics
  • Cell Biology

Background:

  • Circulating microparticles (MPs) are physiological and change in disease.
  • The normal MP proteome lacks characterization using standardized high-resolution methods.

Purpose of the Study:

  • To quantitatively profile the normal microparticle proteome using standardized high-resolution methods.
  • To establish a reproducible method for MP preparation and analysis.
  • To provide a baseline MP proteome for future research.

Main Methods:

  • Quantitative proteomic profiling using nano-LC-MS/MS on an LTQ-Orbitrap.
  • Optimized sample collection, preparation, and analysis of 12 normal samples.
  • Label-free quantitation validated by cytoskeletal protein intensities and flow cytometry.

Main Results:

  • Quantified 536 unique proteins, with 334 (63%) forming a core MP proteome present in all samples.
  • Demonstrated <10% variation in technical triplicates within a 5-decade dynamic range.
  • Developed normalization strategies for variable MP numbers and preparation losses using cytoskeletal proteins.

Conclusions:

  • Established a reproducible LC-MS/MS procedure and a robust MP preparation method.
  • Generated a baseline normal MP proteome for comparative studies in health and disease.
  • Validated the utility of cytoskeletal proteins for normalizing MP quantification.