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 language | English |
|---|---|
| Pages (from-to) | 227-236 |
| Number of pages | 10 |
| Journal | Journal of Peptide Science |
| Volume | 13 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Apr 2007 |
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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 journal › Article
}
TY - JOUR
T1 - Mass spectrometric identification of the trypsin cleavage pathway in lysyl-proline containing oligotuftsin peptides
AU - Manea, Marilena
AU - Mező, G.
AU - Hudecz, F.
AU - Przybylski, Michael
PY - 2007/4
Y1 - 2007/4
N2 - 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.
AB - 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.
KW - -Lys-Pro- cleavage
KW - High-resolution FTICR mass spectrometry
KW - Oligotuftsin derivatives
KW - Trypsin specificity
UR - http://www.scopus.com/inward/record.url?scp=34247330927&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34247330927&partnerID=8YFLogxK
U2 - 10.1002/psc.836
DO - 10.1002/psc.836
M3 - Article
C2 - 17394121
AN - SCOPUS:34247330927
VL - 13
SP - 227
EP - 236
JO - Journal of Peptide Science
JF - Journal of Peptide Science
SN - 1075-2617
IS - 4
ER -