Complementary peptide sequence coverage using alternative enzymes for on-line digestion with a triaxial electrospray probe

J Am Soc Mass Spectrom. 2009 Nov;20(11):1983-7. doi: 10.1016/j.jasms.2009.07.022. Epub 2009 Aug 12.

Abstract

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 Abeta fibrils are suggestive of unique reactive intermediates during dissolution. Data obtained with enzyme mixtures suggest synergistic effects.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Amyloid beta-Peptides / chemistry*
  • Animals
  • Aspartic Acid Endopeptidases / chemistry*
  • Hydrogen Bonding
  • Hydrolysis
  • Molecular Sequence Data
  • Online Systems*
  • Pepsin A / metabolism
  • Peptide Fragments / chemistry*
  • Peptide Hydrolases / chemistry*
  • Peptide Hydrolases / classification
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Swine

Substances

  • Amyloid beta-Peptides
  • Peptide Fragments
  • amyloid beta-protein (1-40)
  • Peptide Hydrolases
  • Aspartic Acid Endopeptidases
  • Endothia aspartic proteinase
  • Pepsin A