GTP对大鼠肝微粒体囊泡Ca2+流出的作用机制。通过荧光能量转移测量囊泡融合。
The mechanism of action of GTP on Ca2+ efflux from rat liver microsomal vesicles. Measurement of vesicle fusion by fluorescence energy transfer.
作者信息
Comerford J G, Dawson A P
机构信息
School of Biological Sciences, University of East Anglia, Norwich U.K.
出版信息
Biochem J. 1988 Jan 1;249(1):89-93. doi: 10.1042/bj2490089.
Abstract
- GTP-promoted fusion between microsomal vesicles was studied by using fluorescence-resonance-energy transfer between the fluorescent membrane probes octadecanoyl-aminofluorescein and octadecyl-rhodamine. 2. The fluorescence increase after GTP addition does not require the presence of ATP, is unaffected by changes in free [Ca2+] in the range 10 microM-1 nM, but requires Mg2+, although higher Mg2+ concentrations are inhibitory. 3. In terms of requirements for poly(ethylene glycol), dependence on GTP concentration and inhibition by high Mg2+ concentrations, there is excellent correlation between rate of increase in fluorescence and rate of GTP-promoted Ca2+ efflux measured under Ca2+ transport conditions. 4. The observations support our previous conclusions that GTP-induced membrane fusion plays a major role in causing GTP-promoted Ca2+ efflux from microsomal vesicles.
摘要
- 通过使用荧光膜探针十八烷酰氨基荧光素和十八烷基罗丹明之间的荧光共振能量转移，研究了GTP促进的微粒体囊泡之间的融合。2. 添加GTP后的荧光增加不需要ATP的存在，在10微摩尔/升至1纳摩尔/升范围内游离[Ca2+]的变化对其无影响，但需要Mg2+，尽管较高的Mg2+浓度具有抑制作用。3. 就对聚乙二醇的需求、对GTP浓度的依赖性以及高Mg2+浓度的抑制作用而言，荧光增加速率与在Ca2+转运条件下测量的GTP促进的Ca2+流出速率之间存在极好的相关性。4. 这些观察结果支持了我们之前的结论，即GTP诱导的膜融合在导致GTP促进的微粒体囊泡Ca2+流出中起主要作用。
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