caa a22 4500 397555202 CHVBK 20180308164759.0 cr unu---uuuuu 161202e199602 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021244 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021244 Purification and Identification of a Novel and Four Known Serine Proteinase Inhibitors Secreted by Human Glioblastoma Cells [Elektronische Daten] [Naohiko Koshikawa, Takashi Nakamura, Naoto Tsuchiya, Mitsuko Isaji, Hidetaro Yasumitsu, Makoto Umeda, Kaoru Miyazaki] Our previous studies have shown that some human cancer cell lines produce pancreatic trypsinogen, plasminogen, and tissue-type kallikrein. To understand the regulatory mechanism of these proteinases, serine proteinase inhibitors secreted by human glioblastoma cell line T98G were analyzed by gelatin reverse zymography with trypsin. The serum-free conditioned medium of T98G cells showed more than ten trypsin inhibitor bands ranging from 16 to 150 kDa in the reverse zymography. Major trypsin inhibitors were purified by trypsin-afflnity chromatography. Analysis of their N-terminal amino acid sequences demonstrated that the purified inhibitors were identical to the secreted forms of amyloid protein precursors (APPs), tissue factor pathway inhibitor (TFPI), placental protein 5 (PP5)/TFPI-2, and secretory leukocyte proteinase inhibitor (SLPI). In addition, a novel 25-kDa trypsin-binding protein, tentatively named p25TI, was identified. p25TI showed weak inhibitory activity against trypsin in reverse zymography as compared with the other inhibitors. The secretion of multiple forms of serine proteinase inhibitors by human cancer cells raises the possibility that they might be involved in the abnormal growth of cancer cells. © by the Journal of Biochemistry Regular Papers nationallicence cancer nationallicence glioblastoma nationallicence reverse zymography nationallicence serine proteinase inhibitor nationallicence trypsin inhibitor nationallicence Koshikawa Naohiko Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut Nakamura Takashi Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut Tsuchiya Naoto Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut Isaji Mitsuko Bioscience Laboratory, Mochida Pharmaceuticals 1-1-1 Kamiya, Kita-ku, Tokyo 115 aut Yasumitsu Hidetaro Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut Umeda Makoto Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut Miyazaki Kaoru Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut The Journal of Biochemistry Oxford University Press 119/2(1996-02), 334-339 0021-924X 119:2<334 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021244 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021244 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Koshikawa Naohiko Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut NATIONALLICENCE 700 1- Nakamura Takashi Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut NATIONALLICENCE 700 1- Tsuchiya Naoto Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut NATIONALLICENCE 700 1- Isaji Mitsuko Bioscience Laboratory, Mochida Pharmaceuticals 1-1-1 Kamiya, Kita-ku, Tokyo 115 aut NATIONALLICENCE 700 1- Yasumitsu Hidetaro Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut NATIONALLICENCE 700 1- Umeda Makoto Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut NATIONALLICENCE 700 1- Miyazaki Kaoru Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University 641-12 Maioka-cho, Totsuka-ku, Yokohama 244 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/2(1996-02), 334-339 0021-924X 119:2<334 1996 119 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