Peptide sequence information derived by pronase digestion and ammonium sulfate in-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

J Am Soc Mass Spectrom. 2000 Nov;11(11):1000-8. doi: 10.1016/S1044-0305(00)00170-7.

Abstract

We present the use of Pronase digestion and in-source decay in the presence of ammonium sulfate as complementary techniques to confirm the amino acid sequence of a peptide. Pronase, a commercial preparation from Streptomyces griseus, is a combination of proteolytic enzymes. It produces carboxypeptidase and aminopeptidase ladders using a single Pronase digestion and represents an inexpensive, nonspecific, and fast supplement to traditional sequencing enzymes. However, N-terminal peptidase activity appears dependent on the terminal amino acid residue. We also introduce the use of saturated ammonium sulfate as an "on-slide" sample additive to promote in-source fragmentation of peptides. Use of saturated ammonium sulfate resulted in a simple way to increase peptide backbone fragmentation and essentially produced either a cn or yn ion series. Together these techniques provide useful supplements to existing methods for peptide sequence information.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adrenocorticotropic Hormone / chemistry
  • Amino Acid Sequence
  • Ammonium Sulfate / chemistry*
  • Bradykinin / chemistry
  • Hydrolysis
  • Indicators and Reagents
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptides / analysis*
  • Peptides / chemistry
  • Pronase / chemistry*
  • Salmonella / chemistry
  • Sequence Analysis
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Indicators and Reagents
  • Peptide Fragments
  • Peptides
  • Adrenocorticotropic Hormone
  • Pronase
  • Bradykinin
  • Ammonium Sulfate