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Related Concept Videos

Proteomics01:33

Proteomics

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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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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
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Proteome Integral Solubility Alteration: A High-Throughput Proteomics Assay for Target Deconvolution.

Massimiliano Gaetani1,2, Pierre Sabatier1, Amir A Saei1

  • 1Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics , Karolinska Institutet , SE-17 177 Stockholm , Sweden.

Journal of Proteome Research
|September 24, 2019
PubMed
Summary

The Proteome Integral Solubility Alteration (PISA) assay enhances protein solubility analysis. This method significantly increases throughput and reduces compound and biological material usage for drug development.

Keywords:
action mechanismchemical biologydrug developmenthigh throughputmass spectrometryprotein solubilityprotein stabilityproteomicstandem mass tagtarget deconvolution

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Drug Discovery

Background:

  • Agents like drugs alter protein physicochemical properties in biological systems.
  • Protein alterations can be measured using stability- and solubility-modifying factors, such as temperature and agent concentration.
  • Traditional methods for analyzing these alterations are often low-throughput.

Purpose of the Study:

  • To introduce the Proteome Integral Solubility Alteration (PISA) assay.
  • To demonstrate PISA's ability to analyze protein solubility changes with high throughput.
  • To highlight PISA's efficiency in reducing sample and compound consumption.

Main Methods:

  • The PISA assay utilizes varying degrees of a stability- and solubility-modifying factor (e.g., temperature) combined with varying agent concentrations.
  • It measures changes in protein solubility across multiple conditions.
  • Analysis is performed without the need for curve fitting to solubility data.

Main Results:

  • PISA increases analysis throughput by 1 to 2 orders of magnitude compared to standard methods.
  • The assay significantly reduces the consumption of compounds and biological materials.
  • It allows for an unlimited number of factor variation points.

Conclusions:

  • The PISA assay offers a highly efficient method for probing protein alterations.
  • This approach is well-suited for applications in chemical biology and drug development.
  • Widespread adoption of PISA is anticipated in these fields.