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Collagen type II modification by hypochlorite.

Sławomir Olszowski1, Paweł Mak, Ewa Olszowska

  • 1Institute of Medical Chemistry, Collegium Medicum, Jagiellonian University, 31-121 Kraków, Czysta 18, Poland.

Acta Biochimica Polonica
|July 2, 2003
PubMed
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Hypochlorite oxidation of collagen type II (CII) in rheumatoid arthritis causes deamination, dichlorotyrosine formation, and fragmentation. Changes in CII aggregate size serve as a key marker for this oxidative damage.

Area of Science:

  • Biochemistry
  • Immunology
  • Rheumatology

Background:

  • Protein oxidation is a key inflammatory process, with hypochlorite (HOCl/OCl(-)) from neutrophils implicated in rheumatoid arthritis.
  • Collagen type II (CII), the main cartilage protein, is a significant target of HOCl/OCl(-) in joint destruction.

Purpose of the Study:

  • To investigate the modifications of collagen type II induced by hypochlorite (HOCl/OCl(-)) and ammonium chloramine (NH(2)Cl).
  • To characterize the structural and aggregate changes in CII following chlorination.
  • To identify potential biomarkers for hypochlorite-mediated collagen oxidation.

Main Methods:

  • Circular dichroism (CD) spectroscopy
  • High-performance liquid chromatography (HPLC)
  • Enzyme-linked immunosorbent assay (ELISA)

Related Experiment Videos

  • Dynamic light scattering (DLS)
  • Fluorimetry
  • Spectrophotometry

    • Main Results:

    • Hypochlorite treatment caused deamination, carbonyl group formation, and tyrosine to dichlorotyrosine conversion in CII.
    • Ammonium chloramine also modified CII, but with 50% lower yield of carbonyl groups and dichlorotyrosine compared to hypochlorite.
    • CD data indicated CII denaturation and fragmentation, with peptide sizes dependent on hypochlorite concentration.
    • DLS revealed a decrease in collagen II aggregate radius from 30 nm to 6.8 nm, suggesting smaller micelle formation.
    • Collagen chlorination altered epitopes, impacting recognition by anti-CII antibodies.

    Conclusions:

    • Hypochlorite significantly modifies collagen type II structure and integrity.
    • The size of molecular aggregates of collagen II is a sensitive marker for hypochlorite-mediated oxidation in inflammatory conditions like rheumatoid arthritis.
    • Understanding these modifications is crucial for developing diagnostic markers for joint destruction in inflammatory diseases.