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首页> 外文期刊>Cell biology international. >GTP-binding proteins G(salpha), G(ialpha), and Ran identified in mitochondria of human placenta.
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GTP-binding proteins G(salpha), G(ialpha), and Ran identified in mitochondria of human placenta.

机译：GTP结合蛋白G（salpha），G（ialpha）和Ran在人胎盘的线粒体中鉴定。

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GTP-binding proteins (GTPases) have been detected in the mitochondria of human placenta. It has been proposed that porin interacts with GTPases in the mitochondrion to modulate contact site function, however, their identity and location is not known. In this study, we investigated the location of GTPases in mitochondria from term placentae as well as the expression of mitochondrial GTPases in mid-term placentae. Mitochondria obtained from human term and mid-term placentae were purified by sedimentation. Sub-mitochondrial vesicles prepared from ruptured and sonicated mitochondria were separated by ultracentrifugation in sucrose density gradients. The location of membrane vesicles was determined using marker enzymes. Mitochondrial proteins were separated by SDS-PAGE. Western blots were incubated in [alpha-(32)P]-GTP and detected using autoradiography or antibodies against known GTPases and porin followed by enhanced chemiluminescence. [alpha-(32)P]-GTP bound 24 and 28 kDa proteins located in the outer membrane. The G(salpha)antibody detected 42.5, 53 and 67 kDa proteins. The G(ialpha)antibody identified a 40.5 kDa band in contact sites and the outer membrane, as well as 55 and 105 kDa proteins in contact site vesicles. The Ran antibody detected a 28 kDa protein, mainly in the outer membrane. Porin migrated at 30 kDa. G(ialpha)and Ran were detected in mitochondria from both term and mid-term placentae. The location of porin and GTPases leave open the possibility that these proteins interact in contact sites and may also be responding to extra-mitochondrial signals. Ran and G(ialpha)are expressed by mid-term in human placentae and may be necessary for placental functions at this stage of development. It will be important in future experiments to characterise the physiological functions of these GTP-binding proteins in the mitochondria of human placenta.
机译：在人胎盘的线粒体中已检测到GTP结合蛋白（GTPase）。已经提出，孔蛋白与线粒体中的GTP酶相互作用以调节接触部位的功能，但是，其身份和位置尚不清楚。在这项研究中，我们调查了足月胎盘中线粒体中GTPases的位置以及中期胎盘中线粒体GTPases的表达。通过沉降纯化从人足和中期胎盘获得的线粒体。通过破裂和超声处理的线粒体制备的线粒体亚囊泡通过蔗糖密度梯度超速离心分离。使用标记酶确定膜囊泡的位置。线粒体蛋白通过SDS-PAGE分离。 Western印迹在[alpha-（32）P] -GTP中孵育，并使用放射自显影或针对已知GTPases和孔蛋白的抗体进行检测，然后增强化学发光。 [α-（32）P] -GTP结合位于外膜的24和28 kDa蛋白。 G（salpha）抗体检测到42.5、53和67 kDa蛋白。 G（ialpha）抗体在接触部位和外膜中鉴定出40.5 kDa的条带，在接触部位囊泡中鉴定出55和105 kDa的蛋白质。 Ran抗体检测到28 kDa蛋白，主要在外膜中。孔蛋白以30 kDa迁移。在足月和中期胎盘的线粒体中均检测到G（ialpha）和Ran。孔蛋白和GTPases的位置使这些蛋白质在接触部位相互作用的可能性敞开，并且还可能对线粒体外信号作出反应。 Ran和G（ialpha）在人胎盘中期表达，在这个发育阶段可能是胎盘功能所必需的。在未来的实验中，表征这些GTP结合蛋白在人胎盘线粒体中的生理功能将非常重要。

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