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Journal of Biochemistry Advance Access originally published online on December 11, 2006
Journal of Biochemistry 2007 141(1):69-76; doi:10.1093/jb/mvm005
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© 2006 The Japanese Biochemical Society.

Quantification of Neutral Cysteine Protease Bleomycin Hydrolase and its Localization in Rat Tissues

Yayoi Kamata1, Yoshiko Itoh2, Akane Kajiya1, Sachiyo Karasawa1, Chie Sakatani1, Susumu Takekoshi2, R. Yoshiyuki Osamura2 and Atsushi Takeda1,*

1Laboratory of Biochemistry, Faculty of Arts and Sciences, Sagami Women's University, Sagamihara, Kanagawa 228-8533; and 2Department of Pathology, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan

*To whom correspondence should be addressed. Laboratory of Biochemistry, Faculty of Arts and Sciences, Sagami Women's University, Sagamihara, Kanagawa 228-8533. Tel: +81-42-742-1428, Fax: +81-42-743-4717, E-mail: takeda.a.ns{at}star.sagami-wu.ac.jp

Received August 10, 2006; Accepted November 13, 2006


  Abstract

A neutral cysteine protease, bleomycin hydrolase (BH), was found to be present in the range 3.7–131.1 ng per mg of rat tissues by enzyme-lined immunosorbent assay (ELISA). Newborn rat skin contained the highest amount of BH, and relatively high levels of BH were detected in the kidney and liver of 6-week-old male rats. The tissue distribution of BH in female rats was similar to that in male rats. Moreover, BH was detected in the extracts of erythrocytes and leukocyte-rich cells as well as in those of rat hemo-lymphocytic lineage cell lines by Western blotting. The BH level was increased at 6 weeks after birth and then slightly decreased. By immunohistochemistry, BH was localized as granular staining in the distal and proximal tubular cells of the kidney, and it was also detected in hepatocytes of the liver, in the red pulpy region of the spleen and in neurons of the brain. An immunoelectron microscopic study showed that BH-immunoreactivity was essentially located in the cytoplasm and at the outer membrane of the rough endoplasmic reticulum of epithelial cells of the kidney, as well as in that of hepatocytes of the liver. These results suggest that BH may play ubiquitous and unique roles in rat tissues.

Key Words: bleomycin hydrolase, cysteine protease, ELISA, immunocytochemistry, rat tissues

Abbreviations: BH, bleomycin hydrolase; Cit-ß-NA, Citrulline-ß-naphthylamide; ELISA, enzyme-linked immunosorbent assay


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Y. Kamata, A. Taniguchi, M. Yamamoto, J. Nomura, K. Ishihara, H. Takahara, T. Hibino, and A. Takeda
Neutral Cysteine Protease Bleomycin Hydrolase Is Essential for the Breakdown of Deiminated Filaggrin into Amino Acids
J. Biol. Chem., May 8, 2009; 284(19): 12829 - 12836.
[Abstract] [Full Text] [PDF]


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