caa a22 4500 397494602 CHVBK 20180308164518.0 cr unu---uuuuu 161202e199510 xx s 000 0 eng 10.1093/jb/118.5.1037 doi (NATIONALLICENCE)oxford-10.1093/jb/118.5.1037 Lysine-49-Phospholipases A2 from Trimeresurus flavoviridis Venom Are Membrane-Acting Enzymes [Elektronische Daten] [Yasuyuki Shimohigashi, Ayako Tani, Hiroshi Matsumoto, Kin-ichi Nakashima, Yoko Yamuguchi, Naoko Oda, Yukio Takano, Hiro-o Kamiya, Junji Kishino, Hitoshi Arita, Motonori Ohno] Basic proteins I and II (BP-I and BP-II) isolated from the venom of Trimeresurus flavoviridis (Habu snake) are isozymes of highly active Asp-49-phospholipase A2 (Asp-49-PLA2) and classified into the group Lys-49-PLA2. BP-II was found to elicit a strong contraction of guinea pig ileum, and this activity was inhibited completely by 1 μM indomethacin, an inhibitor of the arachidonate cascade. BP-II was inactive in the Ca2+ -free medium, and p-bromophenacy lated His-48-BP-II was also inactive. BP-II exhibited no binding affinity for the cells expressinng PLA2 receptors. These results indicated that the contraction elicited by BP-II is due to the hydrolytic action of BP-II, liberating archidonic acid from the ileum phospholipid biomembranes. In spite of its limited lipolytic activities (av. 0.9% of Asp-49-PLA2) against monomers and micelles of synthetic phospholipids, BP-II hydrolyzed considerably strongly the phospholipids in the artificial bilayer vesicles. Arachidonic acid released from liposomes of β-arachidonoyl-γ-stearoyl-l-α-phosphatidylcholic was determined by HPLC, and the activity of BP-II was estimated to be about 75% as compared to Asp--49-PLA2. Liposomes encapsulating carboxyflurescein exhibited a strong dye-leakage induced by BP-II in a concentration-dependent manner, only in the Ca2+-containing buffer. The net result from all these observations was that BP-II, a Lys-49-PLA2, is an enzyme that hyrolyzes the membrane phospholipids. In contrast to BP-II, BP-I was found to be considerably weak in hydrolyzing membrane phospholipids, although its activities were distinct. BP-I and BP-Ii share a common sequence with the sole exception of Asp-67 (BP-I) and Asn-67 (BP-II) in the aligned sequences. This implies that the amino acid at position 67 of Lys-49-PLA2s is the residue required for discriminatory recognition of β-arachidonoyl-phospholipid membranes. © Oxford University Press Regular Papers nationallicence isozymes nationallicence lipolytic activity nationallicence liposomes nationallicence Lys-49-phospholipase A2 nationallicence phospholipase A2 nationallicence Shimohigashi Yasuyuki Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut Tani Ayako Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut Matsumoto Hiroshi Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut Nakashima Kin-ichi Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut Yamuguchi Yoko Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut Oda Naoko Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut Takano Yukio Department of Pharmcology, Faculty of Pharmaceutical Sciences, Fukuoka University Fukuoka 814-80 aut Kamiya Hiro-o Department of Pharmcology, Faculty of Pharmaceutical Sciences, Fukuoka University Fukuoka 814-80 aut Kishino Junji Shionogi Research Laboratories, Shionogi & Co., Ltd., Fukushima-ku, Osaka 553 aut Arita Hitoshi Shionogi Research Laboratories, Shionogi & Co., Ltd., Fukushima-ku, Osaka 553 aut Ohno Motonori Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut The Journal of Biochemistry Oxford University Press 118/5(1995-10), 1037-1044 0021-924X 118:5<1037 1995 118 jbchem https://doi.org/10.1093/jb/118.5.1037 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/jb/118.5.1037 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Shimohigashi Yasuyuki Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Tani Ayako Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Matsumoto Hiroshi Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Nakashima Kin-ichi Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Yamuguchi Yoko Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Oda Naoko Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 700 1- Takano Yukio Department of Pharmcology, Faculty of Pharmaceutical Sciences, Fukuoka University Fukuoka 814-80 aut NATIONALLICENCE 700 1- Kamiya Hiro-o Department of Pharmcology, Faculty of Pharmaceutical Sciences, Fukuoka University Fukuoka 814-80 aut NATIONALLICENCE 700 1- Kishino Junji Shionogi Research Laboratories, Shionogi & Co., Ltd., Fukushima-ku, Osaka 553 aut NATIONALLICENCE 700 1- Arita Hitoshi Shionogi Research Laboratories, Shionogi & Co., Ltd., Fukushima-ku, Osaka 553 aut NATIONALLICENCE 700 1- Ohno Motonori Laboratory of Biochemistry, Deparment of Chemistry, Faculty of Science, Kyushu University Fukuoka 812-81 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 118/5(1995-10), 1037-1044 0021-924X 118:5<1037 1995 118 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