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식물세포에서 분리한 Tonoplast Vesicle에서의 $Ca^{2+}$ 수송과 Inositol 1, 4, 5-trisphosphate $(IP_3)$의 작용
김현주,이준승,Kim, Hyeon-Joo,Lee, June-Seung 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.4
오옥신의 작용이 $Ca^{2+}$을 매개로 일어나는지를 확인하기 위하여 Red Beet Root (Beta vulgaris L.)에서 Metrizamide discontinuous density-gradient centrifugation 방법으로 $Ca^{2+}$ pool 이라고 생각되는 tonoplast vesicle을 분리하여 오옥신이 tonoplast vesicle로부터의 $Ca^{2+}$방출에 어떤 영향을 미치는가를 조사하였다. 분리된 tonoplast vesicle내로 $^{45}Ca^{2+}$은 ATP dependent한 과정에 의해 흡입되며 이러한 tonoplast vesicle내로의 $^{45}Ca^{2+}$ 유출입은 Diltiazem, Verapamil, Flunarizine 같은 calcium entry blocker에 의해 억제된다. 오옥신은 $^{45}Ca^{2+}$이 미리 loading된 tonoplast vesicle로부터의 $^{45}Ca^{2+}$방출에 직접적인 영향을 미치지 않으나, Inositol 1,4,5-trisphosphate $(IP_3)$는 $^{45}Ca^{2+}$이 ATP dependent한 과정에 의해 미리 loading된 tonoplast vesicle로부터 $^{45}Ca^{2+}$방출을 촉진시키며 $IP_3$에 의한 $Ca^{2+}$ 방출은 $IP_3$농도에 의존한다. 또한 $IP_3$에 의한 tonoplast vesicle로부터의 $^{45}Ca^{2+}$방출은 $Ca^{2+}$ antagonist인 8-(N, N-Diethylamino)-octyl 3,4,5-Trimethoxybenzoate-HCl(TMB-8)에 의해 억제된다. 오옥신이 빙카(Catharanthus roseus)의 $G_1$단계에서 멈춘 세포에서 Phosphatidylinositol의 turnover에 영향을 미친다는 보고(Ettlinger et al., 1988)와 이 실험결과로 오옥신은 세포질의 $Ca^{2+}$농도를 직접 조절하는 것이 아니라 $IP_3$를 Secondary messenger로 하여 세포절의 $Ca^{2+}$농도를 조절한다고 생각된다. Auxin may exert its effects through $Ca^{2+}$ as a secondary messenger. To test this hypothesis, effects of auxin on $Ca^{2+}$ release from isolated tonoplast vesicles was investigated. Tonoplast vesicles were isolated from red beet roots(Beta vulgaris L.) by metrizamide discontinuous density-gradient centrifugation. Uptake of $^{45}Ca^{2+}$ into tonoplast vesicles was ATP-dependent, and was blocked by calcium entry blockers such as Diltiazem, Verapamil, Flunarizin. Auxin did not induce the release of $^{45}Ca^{2+}$ from tonoplast vesicles preloaded with the cations, but Inositol 1,4,5-trisphosphate $(IP_3)$ did. The $IP_3$ induced $^{45}Ca^{2+}$ release was dependent on $IP_3$ concentration, and was blocked by the $Ca^{2+}$ antagonist, 8-(N,N-diethylamino)-octyl 3,4,5-trimethoxybenzoate-HCl (TMB-8). Auxin is known to generate transient change in levels of Inositol 1,4,5-trisphosphate $(IP_3)$ and Inositol bis-phosphate (ptdIns(4,5)$P_2$) within minutes in Catharanthus roseus (Ettlinger, et al., 1988). These results indicate that auxin cannot alter cytoplasmic calcium levels directly but through the $IP_3$ secondary messenger system.
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식물세포에서 분리한 Tonoplast Vesicle 에서의 Ca2+ 수송과 Inositol 1 , 4 , 5 - trisphosphate ( IP 3 ) 의 작용
김현주,이준승 ( Hyeon Joo Kim,June Seung Lee ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.4
Auxin may exert its effects through Ca^(2+) as a secondary messenger. To test this hypothesis, effects of auxin on Ca^(2+) release from isolated tonoplast vesicles was investigated. Tonoplast vesicles were isolated from red beet roots(Beta vulgaris L.) by metrizamide discontinuous density-gradient centrifugation. Uptake of ^(45)Ca^(2+) into tonoplast vesicles was ATP-dependent, and was blocked by calcium entry blockers such as Diltiazem, Verapamil, Flunarizin. Auxin did not induce the release of ^(45)Ca^(2+) from tonoplast vesicles preloaded with the cations, but Inositol 1,4,5-trisphosphate (IP₃) did. The IP₃ induced ^(45)Ca^(2+) release was dependent on IP₃ concentration, and was blocked by the Ca^(2+) antagonist, 8-(N,N-diethylamino)-octyl 3,4,5-trimethoxybenzoate-HCl (TMB-8). Auxin is known to generate transient change in levels of Inositol 1,4,5-trisphosphate (IP₃) and Inositol bis-phosphate (ptdIns(4,5)P₂) within minutes in Catharanthus roseus (Ettlinger, et al., 1988). These results indicate that auxin cannot alter cytoplasmic calcium levels directly but through the IP₃ secondary messenger system.
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옥수수 자엽초에서 오옥신 유발 에틸렌 생성에 대한 Mannose 의 억제작용
조성혜(Seong Hye Cho),김현조(Hyeon Joo Kim),이준승(June Seung Lee) 한국식물학회 1990 Journal of Plant Biology Vol.33 No.4
Effect of mannose on auxin-induced ethylene production in corn (Zea mays L.) coleoptiles was studied. Auxin induced ethylene production decreased in proportion to mannose concentrations. The inhibitory effect of mannose appeared after 2 h of incubation. Ethylene production was significantly depressed by mannose at high concentrations (10^-5 M-10^-4 M) of indole acetic acid (IAA), but not at low concentrations (10^-8 M-10^-6M). The inhibition of auxin-induced ethylene production by mannose was specific, since other sugars such as galactose, glucose, sucrose and mannitol did not have an inhibitory effect. In an effort to elucidate mechanisms of mannose the effect on the auxin induced ethylene production, effect of the sugar on ACC synthase activity and ACC induced ethylene production was studied. Mannose failed to inhibit ACC mediated ethylene production, but decreased both the ACC content and ACC synthase activity in the tissue. These results suggest that the inhibitory effect of mannose on auxin induced ethylene production results from suppression of auxin induction of ACC synthase.
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