Preparation of the Low Molecular Weight Serum Proteome for Mass Spectrometry Analysis

Veenstra, Timothy D.

doi:10.1007/978-1-4939-9597-4_5

Cited by 1 publication

(2 citation statements)

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“…12 Veenstra used a method to analyze serum peptides after they were separated from large proteins. 13 Differential protein precipitation methods can be useful as a strategy for fractionating proteins. Merrihew et al used ammonium sulfate precipitation to fractionate C. elegans' proteins, which increased coverage of the proteome to validate open reading frames predicted in the genome.…”

Section: ■ Introductionmentioning

confidence: 99%

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Differential Precipitation of Proteins: A Simple Protein Fractionation Strategy to Gain Biological Insights with Proteomics

Pinto,

Diedrich,

Moresco

et al. 2023

J. Am. Soc. Mass Spectrom.

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Differential precipitation of proteins (DiffPOP) is a simple technique for fractionating complex protein mixtures. Using stepwise addition of acidified methanol, ten distinct subsets of proteins can be selectively precipitated by centrifugation and identified by mass spectrometry-based proteomics. We have previously shown that the ability of a protein to resist precipitation can be altered by drug binding, which enabled us to identify a novel drug−target interaction. Here, we show that the addition of DiffPOP to a standard LC−MS proteomics workflow results in a three-dimensional separation of peptides that increases protein coverage and peptide identifications. Importantly, DiffPOP reveals solubility differences between proteoforms, potentially providing valuable insights that are typically lost in bottom-up proteomics.

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Section: ■ Introductionmentioning

confidence: 99%

“…Differential fractionation methods are a traditional biochemical method to enrich proteins or other types of molecules . Veenstra used a method to analyze serum peptides after they were separated from large proteins . Differential protein precipitation methods can be useful as a strategy for fractionating proteins.…”

Section: Introductionmentioning

confidence: 99%