A triaxial probe for on-line proteolysis coupled with hydrogen/deuterium exchange-electrospray mass spectrometry.
An on-line proteolysis system utilizing a triaxial electrospray probe was developed to aid localization of the hydrogen-bonding interaction sites in hydrogen/deuterium exchange-mass spectrometry (HDX-MS) studies of Abeta (1-40) fibrils. The probe allows delayed introduction of the organic solvent component needed for stable electrospray, thus enhancing hydrolysis performance relative to that of a coaxial probe. Effective on-line digestion was accomplished in approximately 12 s. The probe should be of general utility for HDX-MS studies of amyloid fibrils and other protein aggregates.
Chen
Maolian
M
Department of Chemistry, University of Tennessee, Knoxville, Tennessee 37996-1600, USA.
Cook
Kelsey D
KD
Kheterpal
Indu
I
Wetzel
Ronald
R
D016229:Amyloid beta-Peptides / Q000737:chemistry*; D000818:Animals; D041961:Deuterium Exchange Measurement; D006859:Hydrogen / Q000737:chemistry*; D006860:Hydrogen Bonding; D006868:Hydrolysis; D010434:Pepsin A / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry*; D010449:Peptide Mapping; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013552:Swine
D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C078592:amyloid beta-protein (1-40); D006859:Hydrogen; D010434:Pepsin A