Mass spectrometric identification of the trypsin cleavage pathway in lysyl-proline containing oligotuftsin peptides

Marilena Manea, G. Mező, F. Hudecz, Michael Przybylski

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Trypsin cleaves specifically peptide bonds at the C-terminal side of lysine and arginine residues, except for -Arg-Pro-and -Lys-Pro- bonds which are normally resistant to proteolysis. Here we report evidence for a -Lys-Pro- tryptic cleavage in modified oligotuftsin derivatives, AC-[TKPKG]4-NH2) (1), using high-resolution mass spectrometry and HPLC as primary methods for analysis of proteolytic reactions. The proteolytic susceptibility of -Lys-Pro- bonds was strongly dependent on flanking residues, and the flexibility of the peptide backbone might be a prerequisite for this unusual cleavage. While -Lys-Gly- bonds in 1 were rapidly cleaved, the modification of these Lys residues by the attachment of a β-amyloid(4-10) epitope to yield -Lys(X)-Gly derivatives prevented cleavage of this bond, and provided trypsin cleavage of -Lys-Pro- bonds, the pathway of this degradation being independent on the type of Lys-Nε-side chains (acetyl group, amino, acid, peptide). Substitution of the Lys residues by Ala at the P2 positions decreased the tryptic cleavage, while replacement of the bulky side chain of Thr at the P2 positions strongly increased the cleavage of -Lys-Pro- bonds. Circular dichroism (CD) data of the modified oligotuftsin derivatives are in accord with enhanced flexibility of the peptide backbone, as a prerequisite for increased susceptibility to cleavage of -Lys-Pro- bonds. These results obtained of oligotuftsin derivatives might have implications for the proteolytic degradation of target peptides that require specific conformational preconditions.

Original languageEnglish
Pages (from-to)227-236
Number of pages10
JournalJournal of Peptide Science
Volume13
Issue number4
DOIs
Publication statusPublished - Apr 2007

Fingerprint

lysylproline
Proline
Trypsin
Peptides
Derivatives
lysylglycine
arginylproline
Proteolysis
Degradation
Amyloid
Lysine
Mass spectrometry
Arginine
Epitopes
Circular Dichroism
Substitution reactions
Mass Spectrometry
Amino Acids
High Pressure Liquid Chromatography

Keywords

  • -Lys-Pro- cleavage
  • High-resolution FTICR mass spectrometry
  • Oligotuftsin derivatives
  • Trypsin specificity

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Mass spectrometric identification of the trypsin cleavage pathway in lysyl-proline containing oligotuftsin peptides. / Manea, Marilena; Mező, G.; Hudecz, F.; Przybylski, Michael.

In: Journal of Peptide Science, Vol. 13, No. 4, 04.2007, p. 227-236.

Research output: Contribution to journalArticle

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