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Journal of Biochemistry Advance Access published online on October 19, 2007
Journal of Biochemistry, doi:10.1093/jb/mvm170
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© 2007 The Japanese Biochemical Society

Immunochemical and immunohistochemical studies on distribution of elastin fibers in human atherosclerotic lesions using a polyclonal antibody to elastin-derived hexapeptide repeat

Iori Maeda1, Sonoko Kishita2, Yuji Yamamoto1, Kazunari Arima2, Keiko Ideta3, Jing Meng4, Noriyuki Sakata4 and Kouji Okamoto1

1Department of Bioscience and Bioinformatics, Kyushu Institute of Technology, 680-4 Kawazu, Iizuka, Fukuoka 820-8502, Japan
2Department of Chemistry and BioScience, Faculty of Science, Kagoshima University 1-21-35 Korimoto, Kagoshima-city, Kagoshima 890-0065, Japan
3Institute for Material Chemistry and Engineering, Kyushu University, Kasuga, Fukuoka 816-8580, Japan
4Second Department of Pathology, Fukuoka University School of Medicine, 7-45-1 Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan

Correspondence to Iori Maeda, Department of Bioscience and Bioinformatics, Kyushu Institute of Technology, 680-4, Kawazu, Iizuka, Fukuoka 820-8502, Japan., TEL: +81-948-29-7830, FAX: +81-948-29-7801, E-mail: iori{at}bio.kyutech.ac.jp. Correspondence to Kouji Okamoto, Department of Bioscience and Bioinformatics, Kyushu Institute of Technology, 680-4, Kawazu, Iizuka, Fukuoka 820-8502, Japan., TEL: +81-948-29-7810, FAX: +81-948-29-7801, E-mail: okagen{at}bio.kyutech.ac.jp

Received July 7, 2007; Accepted August 28, 2007


  Abstract

A polyclonal antibody to elastin-derived hexapeptide repeat, H-(Val-Gly-Val-Ala-Pro-Gly)3-NH2, was prepared in order to investigate the differences between elastin fibers in intimal hyperplasia and media in human atheroscleroic lesions. The hexapeptide repeat and {alpha}-elastin were recognized by this polyclonal antibody in ELISA, but other elastin-derived peptides such as tetrapeptide repeat, pentapeptide repeat, and nonapeptide were not. In the series of hexapeptide repeats, H-(VGVAPG)n-NH2 where n is 1 to 7, the polyclonal antibody reacted strongly with oligomers (n=3-7) and weakly with dimer (n=2), but not with monomer (n=1). CD measurements suggested that the ß-turn structure is important for recognition by the polyclonal antibody. In an immunohistochemical study, elastin was stained more strongly in intimal hyperplasia than in media, suggesting that newly synthesized elastin in intimal hyperplasia is morphologically distinct from that in media.

Key Words: circular dichroism, elastin, immunochemistry, immunohistochemistry, polyclonal antibody


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