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
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Alkanesulfonates / chemistry*
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Angiotensin II / chemistry
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Angiotensin II / metabolism
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Arginine / analogs & derivatives*
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Arginine / chemistry
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Arginine / metabolism
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Bacteriorhodopsins / chemistry
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Bacteriorhodopsins / metabolism
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Chromatography, High Pressure Liquid
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Chymotrypsin / metabolism
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Dioxolanes / chemistry*
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Endopeptidases / metabolism*
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Hydrogen-Ion Concentration
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Hydrophobic and Hydrophilic Interactions
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Membrane Proteins / chemistry
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Membrane Proteins / metabolism
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Molecular Structure
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Muramidase / chemistry
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Muramidase / metabolism
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Myoglobin / chemistry
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Myoglobin / metabolism
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Neurotensin / chemistry
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Neurotensin / metabolism
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Ovalbumin / chemistry
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Ovalbumin / metabolism
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Proteins / chemistry
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Proteins / metabolism*
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Sodium Dodecyl Sulfate / chemistry
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Solubility
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Spectrometry, Mass, Electrospray Ionization
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
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Surface-Active Agents / chemistry
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Trypsin / metabolism
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Ubiquitin / chemistry
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Ubiquitin / metabolism
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Urea / chemistry
Substances
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Alkanesulfonates
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Dioxolanes
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Membrane Proteins
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Myoglobin
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Proteins
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Surface-Active Agents
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Ubiquitin
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sodium 4-((2-methyl-2-undecyl-1,3-dioxolan-4-yl)methoxyl)-1-propanesulfonate
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Angiotensin II
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Sodium Dodecyl Sulfate
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Neurotensin
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Bacteriorhodopsins
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Urea
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Ovalbumin
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Arginine
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benzoylarginine ethyl ester
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Muramidase
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Endopeptidases
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Chymotrypsin
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Trypsin