Enzyme-friendly, mass spectrometry-compatible surfactant for in-solution enzymatic digestion of proteins.
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.
Yu
Ying-Qing
YQ
Life Sciences Research and Development, Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757, USA.
Gilar
Martin
M
Lee
Peter J
PJ
Bouvier
Edouard S P
ES
Gebler
John C
JC
D000476:Alkanesulfonates / Q000737:chemistry*; D000804:Angiotensin II / Q000737:chemistry / Q000378:metabolism; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D001436:Bacteriorhodopsins / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000378:metabolism; D004148:Dioxolanes / Q000737:chemistry*; D010450:Endopeptidases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry / Q000378:metabolism; D009496:Neurotensin / Q000737:chemistry / Q000378:metabolism; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D012995:Solubility; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013501:Surface-Active Agents / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism; D014508:Urea / Q000737:chemistry
D000476:Alkanesulfonates; D004148:Dioxolanes; D008565:Membrane Proteins; D009211:Myoglobin; D011506:Proteins; D013501:Surface-Active Agents; D025801:Ubiquitin; C472187:sodium 4-((2-methyl-2-undecyl-1,3-dioxolan-4-yl)methoxyl)-1-propanesulfonate; D000804:Angiotensin II; D012967:Sodium Dodecyl Sulfate; D009496:Neurotensin; D001436:Bacteriorhodopsins; D014508:Urea; D010047:Ovalbumin; D001120:Arginine; C009727:benzoylarginine ethyl ester; D009113:Muramidase; D010450:Endopeptidases; D002918:Chymotrypsin; D014357:Trypsin