Selective Site-Specific Fenton Oxidation of Methionine in Model Peptides: Evidence for a Metal-Bound Oxidant

Dawn R. Dufield, George S. Wilson, Richard S Glass, Christian Schöneich

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The metal-catalyzed oxidation (MCO) of proteins represents an important pathway for protein degradation. Although many mechanistic details of MCO are currently unknown, such mechanistic information would greatly benefit formulation scientists in the rational design and analysis of protein formulations. Here, we describe the Fenton oxidation (by Fe2+/H 2O2) of several Met-, Tyr-, and His containing model peptides, including one derivative containing a conformationally restricted norbornyl Met analogue (Nor), Nor-Gly-His-Met-NH2. Our results will provide evidence for a metal-bound reactive oxygen species selectively oxidizing Met to Met sulfoxide, indicating a Met-specific oxidant and arguing against the involvement of freely diffusible hydroxyl radicals. The Fenton oxidation of Nor-Gly-His-Met-NH2 yields a 2:1 preference for sulfoxide formation at the C-terminal Met versus the N-terminal Nor residue, respectively, while incubation of the peptide with H2O2 alone results in a 1:1 ratio. These results are rationalized by the better access of the thioether side chain of the flexible C-terminal Met residue to the peptide-bound iron compared with the conformationally restricted Nor residue. It is commonly believed that Fenton oxidation reactions involve hydroxyl radicals, and that Met oxidation in proteins is predominantly controlled by the surface-accessibility of the respective Met residues. However, occasionally protein oxidation in formulations shows selectivities, which are not consistent with these paradigms. Our results demonstrate additional features of the Fenton reaction such as the formation of a metal-bound oxidant specific for Met (and not Tyr or His), which may assist formulation scientists in the rationalization of unexpected oxidation selectivities.

Original languageEnglish (US)
Pages (from-to)1122-1130
Number of pages9
JournalJournal of Pharmaceutical Sciences
Volume93
Issue number5
DOIs
StatePublished - May 2004

Fingerprint

sulfoxide
Oxidants
Methionine
Metals
Oxidation
Peptides
Hydroxyl Radical
Proteins
Sulfides
Proteolysis
Reactive Oxygen Species
Iron
Derivatives
Degradation

Keywords

  • Fenton reaction
  • Met
  • Met sulfoxide
  • Reactive oxygen species

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Chemistry(all)
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science

Cite this

Selective Site-Specific Fenton Oxidation of Methionine in Model Peptides : Evidence for a Metal-Bound Oxidant. / Dufield, Dawn R.; Wilson, George S.; Glass, Richard S; Schöneich, Christian.

In: Journal of Pharmaceutical Sciences, Vol. 93, No. 5, 05.2004, p. 1122-1130.

Research output: Contribution to journalArticle

Dufield, Dawn R. ; Wilson, George S. ; Glass, Richard S ; Schöneich, Christian. / Selective Site-Specific Fenton Oxidation of Methionine in Model Peptides : Evidence for a Metal-Bound Oxidant. In: Journal of Pharmaceutical Sciences. 2004 ; Vol. 93, No. 5. pp. 1122-1130.
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