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

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Journal of the American Society for Mass Spectrometry

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 ym ion series. Together these techniques provide useful supplements to existing methods for peptide sequence information.

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Authors and Affiliations

  1. Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

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

Authors
  1. Lisa A. Marzilli
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  2. Tamara R. Golden
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  3. Robert J. Cotter
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  4. Amina S. Woods
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Correspondence to Amina S. Woods.

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Marzilli, L.A., Golden, T.R., Cotter, R.J. et al. 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. Spectrom. 11, 1000–1008 (2000). https://doi.org/10.1016/S1044-0305(00)00170-7

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  • DOI: https://doi.org/10.1016/S1044-0305(00)00170-7

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