Enzyme-friendly, mass spectrometry-compatible surfactant for in-solution enzymatic digestion of proteins

Anal Chem. 2003 Nov 1;75(21):6023-8. doi: 10.1021/ac0346196.

Abstract

Improved in-solution tryptic digestion of proteins in terms of speed and peptide coverage was achieved with the aid of a novel acid-labile anionic surfactant (ALS). Unlike SDS, ALS solubilizes proteins without inhibiting trypsin or other common endopeptidases activity. Trypsin activity was evaluated in the presence of various denaturants; little or no decrease in proteolytic activity was observed in 0.1-1% ALS solutions (w/v). Sample preparation prior to mass spectrometry and liquid chromatography analysis consists of sample acidification. ALS degrades rapidly at low-pH conditions, which eliminates surfactant-caused interference with analysis. Described methodology combines the advantages of protein solubilization, rapid digestion, high peptide coverages, and easy sample preparation for mass spectrometry and liquid chromatography analyses.

MeSH terms

  • Alkanesulfonates / chemistry*
  • Angiotensin II / chemistry
  • Angiotensin II / metabolism
  • Arginine / analogs & derivatives*
  • Arginine / chemistry
  • Arginine / metabolism
  • Bacteriorhodopsins / chemistry
  • Bacteriorhodopsins / metabolism
  • Chromatography, High Pressure Liquid
  • Chymotrypsin / metabolism
  • Dioxolanes / chemistry*
  • Endopeptidases / metabolism*
  • Hydrogen-Ion Concentration
  • Hydrophobic and Hydrophilic Interactions
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism
  • Molecular Structure
  • Muramidase / chemistry
  • Muramidase / metabolism
  • Myoglobin / chemistry
  • Myoglobin / metabolism
  • Neurotensin / chemistry
  • Neurotensin / metabolism
  • Ovalbumin / chemistry
  • Ovalbumin / metabolism
  • Proteins / chemistry
  • Proteins / metabolism*
  • Sodium Dodecyl Sulfate / chemistry
  • Solubility
  • Spectrometry, Mass, Electrospray Ionization
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
  • Surface-Active Agents / chemistry
  • Trypsin / metabolism
  • Ubiquitin / chemistry
  • Ubiquitin / metabolism
  • Urea / chemistry

Substances

  • Alkanesulfonates
  • Dioxolanes
  • Membrane Proteins
  • Myoglobin
  • Proteins
  • Surface-Active Agents
  • Ubiquitin
  • sodium 4-((2-methyl-2-undecyl-1,3-dioxolan-4-yl)methoxyl)-1-propanesulfonate
  • Angiotensin II
  • Sodium Dodecyl Sulfate
  • Neurotensin
  • Bacteriorhodopsins
  • Urea
  • Ovalbumin
  • Arginine
  • benzoylarginine ethyl ester
  • Muramidase
  • Endopeptidases
  • Chymotrypsin
  • Trypsin