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蛇毒蛋白Rhodostomin與血小板之交互作用

Rhodostomin (also called kistrin), which was purified from Malayan pit viper (Calloselasma rhodostoma), is a potent anticoagulant. Most anticoagulants identified so far appeared to be the antagonists of fibrinogen receptors, integrin aIIbb3 (GPIIb/IIIa), which mediated activation and biological functions of platelets (Teng and Huang, 1991). They were termed "disintegrin family" for their inhibitory activities on integrin functions and fibrinogen-mediated platelet aggregation (Gould et al., 1990).

Platelets play key roles in many important physiological functions such as hemostasis and thrombosis. Under physiological conditions, platelets need to be activated by a variety of extracellular molecules released from damaged cells, such as ADP or thrombin before fibrinogen can induce cell transformation. However, platelet activation and transformation events were observed when the blood cells interacted with GST-rhodostomin treated substrates, which was, to our knowledge, the first case in which a disintegrin could mediate both activation and morphogenesis effects. These cell transformation events seemed to involve calcium influx and protein phosphorylation. In the demonstration below, I present the calcium spiking of platelets after they attach to the rhodostomin-coated substrate.

 

The calcium spiking observed when platelets were attached to the substrate coated with rhodostomin (left) and the pseudo color applied to the fluorescent signal (right). Before being plated to the substrate, the cells were treated with Calcium Green-1 as a result that the fluorescent signal is proportional to the intracellular concentration of calcium ion. Here we can observe the spreading of the calcium ion from the internal storage of the cell. (Collaborated with Yong-Shyang Yi in Dr. Chi-Hung Lin's lab.)

 

 

The calcium spiking of a platelet attached to the rhodostomin-coated substrate observed by Side View Technology. The cell was trapped and plated precisely by the optical tweezers and simultaneously observed by the Side View Technology. This combined technologies allows us to study the spreading of calcium ion in the initiation of the plate activation on the z-axis. (Collaborated with Yong-Shyang Yi in Dr. Chi-Hung Lin's lab.)

 

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Updated 6/13/2013. Copyright© 2001 Jin-Wu Tsai. All rights reserved.