Complementary Peptide Sequence Coverage Using Alternative Enzymes for On-Line Digestion with a Triaxial Electrospray Probe

Using alternative enzymes for on-line digestion with a triaxial electrospray probe extends sequence coverage. This is the first report of utilization of our triaxial probe for on-line analysis with enzymes other than pepsin, suggesting potential for broader application. The probe allows access to pr...

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Bibliographic Details
Published in:Journal of the American Society for Mass Spectrometry 2009-11, Vol.20 (11), p.1983-1987
Main Authors: Dykstra, Andrew B., Chen, Maolian, Cook, Kelsey D.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amyloid beta-Peptides - chemistry
Analytical Chemistry
Analytical, structural and metabolic biochemistry
Animals
Application Note
Aspartic Acid Endopeptidases - chemistry
Bioinformatics
Biological and medical sciences
Biotechnology
Chemistry
Chemistry and Materials Science
Digestion
Dissolution
Electrospraying
Enzymes
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Hydrogen Bonding
Hydrolysis
Mass spectrometry
Molecular Sequence Data
On-line systems
Online Systems
Organic Chemistry
Pepsin
Pepsin A - metabolism
Peptide Fragments - chemistry
Peptide Hydrolases - chemistry
Peptide Hydrolases - classification
Proteins
Proteomics
Spectrometry, Mass, Electrospray Ionization - methods
Swine
Synergistic effect
Utilization
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Summary:Using alternative enzymes for on-line digestion with a triaxial electrospray probe extends sequence coverage. This is the first report of utilization of our triaxial probe for on-line analysis with enzymes other than pepsin, suggesting potential for broader application. The probe allows access to processes occurring on a timescale and/or involving substrate conformations complementary to those for conventional (off-line) digestion. Some of the features observed in application to Aβ fibrils are suggestive of unique reactive intermediates during dissolution. Data obtained with enzyme mixtures suggest synergistic effects. On-line mixing enables dissolution and hydrolysis on a time-scale that minimizes artifactual exchange. Applicability with multiple enzymes is demonstrated, revealing synergies and possible conformational effects.
ISSN:1044-0305
1879-1123
DOI:10.1016/j.jasms.2009.07.022