caa a22 4500 397556691 CHVBK 20180308164804.0 cr unu---uuuuu 161202e199608 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021420 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021420 Cloning and Analysis of cDNA Encoding Rat Bleomycin Hydrolase, a DNA-Binding Cysteine Protease [Elektronische Daten] [Atsushi Takeda, Yutaka Masuda, Takako Yamamoto, Takahiro Hirabayashi, Yoshiko Nakamura, Kazuyasu Nakaya] We isolated and characterized almost the entire cDNA encoding BLM hydrolase from rat spleen cDNA libraries. The cDNA encoded a polypeptide composed of 454 amino acids, that had a slightly larger molecular mass than that was previously estimated by SDS-PAGE for purified BLhl hydrolase subunit. Amino acid sequence alignments showed that rat BLM hydrolase is homologous to that of rabbit (94% identity of partial amino acid sequence), yeast cysteine protease (39% identity), and the pepC gene products of three bacteria (34-40% identity). In addition, it contained the three regions that are conserved in other cysteine proteases and thought to constitute the catalytic center. These results indicated that rat BLM hydrolase is a member of the papain superfamily of cysteine proteases. Sequencing revealed several putative sites phosphorylated by different types of protein kinases, but no signal sequence, transmembrane domain, N-linked glycosylation site or DNA-binding motif. The yeast homolog is a DNA-binding cysteine protease [Xu et al. (1994) J. Biol. Chem. 269,21177-21183. We demonstrated that rat BLM hydrolase also binds the singlestranded form of the Gal4 DNA-binding site oligonucleotide with high affinity compared with that of the double-stranded form. Northern blots revealed that the level of BLM hydrolase mRNA expression was very low in the rat skin, lung, and skeletal muscle. Furthermore, BLM hydrolase mRNA was ubiquitously expressed in the human cell lines, HeLa, SKG-IIIa, FL, KB, HEp-2, U373 GM, P3HR-1, Raji, THE-1, Jurkat, and Molt-4. These results suggested that BLM hydrolase plays important physiological roles, including the metabolism of antibiotics. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence amino acid sequence nationallicence BLM hydrolase nationallicence cDNA cloning nationallicence cysteine protease nationallicence DNA binding protein nationallicence Takeda Atsushi Department of Clinical Pathology, Showa University Fujigaoka Hospital 1-30 Fujigaoka, Aoba-ku, Yokohama 227 aut Masuda Yutaka Department of Biochemistry, School of Pharmaceutical Sciences, Showa Universiry 1-5-8 Hatanodai Shinagawa-ku, Tokyo 142 aut Yamamoto Takako Department of Clinical Pathology, Showa University Fujigaoka Hospital 1-30 Fujigaoka, Aoba-ku, Yokohama 227 aut Hirabayashi Takahiro Department of Biochemistry, School of Pharmaceutical Sciences, Showa Universiry 1-5-8 Hatanodai Shinagawa-ku, Tokyo 142 aut Nakamura Yoshiko Department of Clinical Pathology, Showa University Fujigaoka Hospital 1-30 Fujigaoka, Aoba-ku, Yokohama 227 aut Nakaya Kazuyasu Department of Biochemistry, School of Pharmaceutical Sciences, Showa Universiry 1-5-8 Hatanodai Shinagawa-ku, Tokyo 142 aut The Journal of Biochemistry Oxford University Press 120/2(1996-08), 353-359 0021-924X 120:2<353 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021420 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021420 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Takeda Atsushi Department of Clinical Pathology, Showa University Fujigaoka Hospital 1-30 Fujigaoka, Aoba-ku, Yokohama 227 aut NATIONALLICENCE 700 1- Masuda Yutaka Department of Biochemistry, School of Pharmaceutical Sciences, Showa Universiry 1-5-8 Hatanodai Shinagawa-ku, Tokyo 142 aut NATIONALLICENCE 700 1- Yamamoto Takako Department of Clinical Pathology, Showa University Fujigaoka Hospital 1-30 Fujigaoka, Aoba-ku, Yokohama 227 aut NATIONALLICENCE 700 1- Hirabayashi Takahiro Department of Biochemistry, School of Pharmaceutical Sciences, Showa Universiry 1-5-8 Hatanodai Shinagawa-ku, Tokyo 142 aut NATIONALLICENCE 700 1- Nakamura Yoshiko Department of Clinical Pathology, Showa University Fujigaoka Hospital 1-30 Fujigaoka, Aoba-ku, Yokohama 227 aut NATIONALLICENCE 700 1- Nakaya Kazuyasu Department of Biochemistry, School of Pharmaceutical Sciences, Showa Universiry 1-5-8 Hatanodai Shinagawa-ku, Tokyo 142 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/2(1996-08), 353-359 0021-924X 120:2<353 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford