caa a22 4500 397555822 CHVBK 20180308164801.0 cr unu---uuuuu 161202e199601 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021217 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021217 Effect of Depolymerized Holothurian Glycosaminoglycan (DHG) on the Activation of Factor VIII and Factor V by Thrombin [Elektronische Daten] [Hideki Nagase, Kei-ichi Enjyoji, Midori Shima, Kenji Kitazato, Akira Yoshioka, Hidehiko Saito, Hisao Kato] Our previous study has shown that depolymerized holothurian glycosaminoglycan (DHG) has two different inhibitory activities in the blood coagulation cascade: heparin cofactor II-dependent thrombin inhibition; and antithrombin III- and heparin cofactor II-indepen-dent inhibition of the intrinsic factor Xase complex [Nagase et al. (1995) Blood 85,15271534]. In the present study, the effect of DHG on the activation of factor VIII and factor V by thrombin was examined with purified human components. DHG inhibited the activation of factor VIII by thrombin at concentrations exceeding 80 nM, but not the activation of factor V by thrombin at concentrations of up to 8 μM. On Western blot analysis, DHG inhibited the cleavage of factor VIII light chain at concentrations exceeding 0.8 μM. The interaction between DHG and factors VIII and V and thrombin was examined with a DHG-cellulofine column. DHG had strong affinity for factor V and thrombin, but slight affinity for factor VIII. The interaction of DHG with thrombin was analyzed, using fluorescein isothiocyan-ate-labeled DHG. One mole of DHG bound 2 mol of thrombin, with a dissociation constant (Kd) of 3.04 ×10−6 M. These results suggest that DHG interferes with the interaction between thrombin and factor VTII, probably by making a binary complex through the anionic binding exosite II of thrombin. © by the Journal of Biochemistry Regular Papers nationallicence anticoagulant nationallicence factor VIII nationallicence glycosaminoglycan nationallicence holothurian nationallicence thrombin nationallicence Nagase Hideki Taiho Pharmaceutical Co., Ltd. 224-2 Ebisuno, Kawauchi-cho, Tokushima 771-01 aut Enjyoji Kei-ichi Research Institute, National Cardiovascular Center 5 Fujishirodai, Suita, Osaka 565 aut Shima Midori the Department of Pediatrics, Nara Medical University Shijyou-machi, Kashihara, Nara 634 aut Kitazato Kenji Taiho Pharmaceutical Co., Ltd. 224-2 Ebisuno, Kawauchi-cho, Tokushima 771-01 aut Yoshioka Akira the Department of Pediatrics, Nara Medical University Shijyou-machi, Kashihara, Nara 634 aut Saito Hidehiko the First Department of Internal Medicine, Nagoya University School of Medicine 65 Tsurumai-cho, Syouwa-ku, Nagoya 466 aut Kato Hisao Research Institute, National Cardiovascular Center 5 Fujishirodai, Suita, Osaka 565 aut The Journal of Biochemistry Oxford University Press 119/1(1996-01), 63-69 0021-924X 119:1<63 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021217 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021217 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nagase Hideki Taiho Pharmaceutical Co., Ltd. 224-2 Ebisuno, Kawauchi-cho, Tokushima 771-01 aut NATIONALLICENCE 700 1- Enjyoji Kei-ichi Research Institute, National Cardiovascular Center 5 Fujishirodai, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Shima Midori the Department of Pediatrics, Nara Medical University Shijyou-machi, Kashihara, Nara 634 aut NATIONALLICENCE 700 1- Kitazato Kenji Taiho Pharmaceutical Co., Ltd. 224-2 Ebisuno, Kawauchi-cho, Tokushima 771-01 aut NATIONALLICENCE 700 1- Yoshioka Akira the Department of Pediatrics, Nara Medical University Shijyou-machi, Kashihara, Nara 634 aut NATIONALLICENCE 700 1- Saito Hidehiko the First Department of Internal Medicine, Nagoya University School of Medicine 65 Tsurumai-cho, Syouwa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Kato Hisao Research Institute, National Cardiovascular Center 5 Fujishirodai, Suita, Osaka 565 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/1(1996-01), 63-69 0021-924X 119:1<63 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