Importance of tryptophan nitration of carbonic anhydrase III for the morbidity of atopic dermatitis.
The nitration of proteins results from the vigorous production of reactive nitrogen species in inflammatory disease. We previously reported the proteomic analysis of nitrated tryptophan residues in in vitro model cells for inflammatory diseases using a 6-nitrotryptophan-specific antibody. In this paper, we applied this method to the analysis of a disease model animal and identified the 6-nitrotryptophan-containing proteins in the skin of atopic dermatitis model mice (AD-NC/Nga mice). We found three nitrotryptophan-containing proteins, namely, carbonic anhydrase III (CAIII), α-enolase (α-ENO), and cytoskeletal keratin type II (KTII), and identified the positions of the nitrotryptophan residues in their amino acid sequences: Trp47 and Trp123 in CAIII, Trp365 in α-ENO, and Trp221 in KTII. Among these, the nitration of CAIII was increased not only in the lesional skin of AD-NC/Nga mice but also in the mice that did not present any symptoms. The in vitro nitration of purified CAIII by peroxynitrite reduced its CO2 hydratase activity in a dose-dependent manner. In addition, we found that CAIII was induced during the differentiation of normal human epidermal keratinocytes. Furthermore, we found the presence of CAIII and the formation of 6-nitrotryptophan-containing proteins in both the lesional and the nonlesional sections of the skin of patients with atopic dermatitis through immunohistochemical staining. This study provides the first demonstration of the formation of 6-nitrotryptophan in human tissues and disease.
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Hiroaki
H
Kawasaki
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Mitsutoshi
M
Tominaga
The Institute of Health and Sports Sciences, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Ayako
A
Shigenaga
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Atsuko
A
Kamo
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Yayoi
Y
Kamata
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Kyoichi
K
Iizumi
Department of Dermatology, Juntendo University Urayasu Hospital, Chiba 279-0021, Japan.
Utako
U
Kimura
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan.
Hideoki
H
Ogawa
The Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Inzai, Chiba 270-1695, Japan; Department of Dermatology, Juntendo University Urayasu Hospital, Chiba 279-0021, Japan.
Kenji
K
Takamori
Juntendo University School of Health Care and Nursing, Chiba 279-0021, Japan. Electronic address: hyamaku@juntendo.ac.jp.
Fumiyuki
F
Yamakura
D000818:Animals; D024403:Carbonic Anhydrase III / Q000378:metabolism*; D002460:Cell Line; D003876:Dermatitis, Atopic / Q000473:pathology*; D004195:Disease Models, Animal; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation / Q000276:immunology / Q000473:pathology; D053557:Keratin-2 / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D030421:Peroxynitrous Acid / Q000737:chemistry; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D051381:Rats; D026361:Reactive Nitrogen Species / Q000737:chemistry; D012867:Skin / Q000473:pathology; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D053557:Keratin-2; D026361:Reactive Nitrogen Species; D030421:Peroxynitrous Acid; C055424:6-nitrotryptophan; D014364:Tryptophan; D024403:Carbonic Anhydrase III; D010751:Phosphopyruvate Hydratase
6-Nitrotryptophan; Atopic dermatitis; Carbonic anhydrase III; Free radicals; NC/Nga mouse; Nitration; Proteomics; Tryptophan residue