The presence of d-β-aspartic acid-containing peptides in elastic fibers of sun-damaged skin: a potent marker for ultraviolet-induced skin aging

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Abstract

Biologically uncommon d-aspartyl residues have been reported in proteins of various elderly tissues. We prepared a polyclonal antibody against d-β-Asp-containing peptide and examined its immunoreactivity in the skin. The antibody recognized integrated or disintegrated elastic fibers in the sun-exposed skin but not in the sun-protected skin of the elderly donors. Western blot analysis of the proteins isolated from sun-damaged skin demonstrated that the 50 kDa protein was immunoreactive with both antibodies for d-β-Asp-containing peptide and elastin. Ultraviolet (UV) irradiation on normal skin caused the appearance of d-β-Asp-containing peptide-immunoreactive fibers in the dermis. These results suggest that UV irradiation is closely related to the formation of d-β-Asp in the elastic fibers of skin. We propose that the antibody could be a useful indicator for sun damage of the skin.

Section snippets

Methods

Antibody againstd-β-Asp-containing peptide. The preparation and characterization of the antibody we used were described in the previous paper [15]. The polyclonal antibody against the peptide; Gly–Leu–d-β-Asp–Ala–Thr–Gly–Leu–d-β-Asp–Ala–Thr–Gly–Leu–d-β-Asp–Ala–Thr (designated peptide 3R), containing three repeats of position 149–153 of the human αA-crystallin optic isomer in rabbit serum, was purified by affinity chromatography using peptide 3R and bovine αA-crystallin as ligands. The antibody

Immunohistochemical staining

The antibody for peptide 3R recognized fibrous network in the upper to mid-dermis of the sun-exposed skin (face) taken from 59-year-old individual (Fig. 1B), but not in the sun-exposed skin (face) of 9-year-old individual (Fig. 1A). The antibody strongly reacted with the elastotic materials of actinic elastosis in the upper dermis of the sun-exposed skin (face) taken from 86-year-old individual (Fig. 1C). In contrast, there was a minimum immunoreactivity of the antibody with fibrous network in

Discussion

We have reported that the configuration of the Asp-151 residue of human αA-crystallin converted to a d-β-Asp isomer during aging [6], [8], [9], [10]. To detect the locality of d-β-Asp-containing αA-crystallin in the lens, we prepared an antibody (anti-peptide 3R antibody) which reacts specifically with d-β-Asp-containing peptides [15]. The antibody crossreacted specifically to d-β-Asp-151-containing αA-crystallin. Immunohistochemical staining of human lens with the antibody demonstrated that d

Acknowledgements

This work was supported by a grant from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

References (21)

  • J.T. Powell et al.

    Atherosclerosis

    (1992)
  • A.E. Roher et al.

    J. Biol. Chem.

    (1993)
  • J.L. Bada

    Methods Enzymol.

    (1984)
  • N. Fujii et al.

    Biochim. Biophys. Acta

    (1994)
  • N. Fujii et al.

    Biophys. Biochem. Res. Commun.

    (1997)
  • N. Fujii et al.

    Biophys. Biochem. Res. Commun.

    (1999)
  • N. Fujii et al.

    Biochem. Biophys. Res. Commun.

    (1999)
  • N. Fujii et al.

    Biochim. Biophys. Acta

    (2001)
  • P.J.T.A. Groenen et al.

    FEBS Lett.

    (1990)
  • N. Fujii et al.

    Exp. Eye Res.

    (1997)
There are more references available in the full text version of this article.

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