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

Lisa A. Marzilli, Tamara R. Golden, Robert J. Cotter, Amina S. Woods

Research output: Contribution to journalArticle

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 Steptomyces 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.

Original languageEnglish (US)
Pages (from-to)1000-1008
Number of pages9
JournalJournal of the American Society for Mass Spectrometry
Volume11
Issue number11
DOIs
StatePublished - Dec 1 2000

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

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