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.
Life Sciences Research and Development, Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757, USA.
Ying-Qing
YQ
Yu
Martin
M
Gilar
Peter J
PJ
Lee
Edouard S P
ES
Bouvier
John C
JC
Gebler
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