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  • Title: Immunoelectron microscopical studies on synthesis and localization of uncoupling protein in brown adipocytes: evidence for cotranslational transport of uncoupling protein into mitochondria.
    Author: Loncar D.
    Journal: J Struct Biol; 1990; 105(1-3):133-45. PubMed ID: 2129216.
    Abstract:
    Through the use of the immunoelectron microscopical technique, uncoupling protein (UCP) was analyzed in the brown adipocytes of room temperature- and cold-acclimated rats, in rat brown adipocytes developed in vitro, and in the brown adipocytes of mice, hamsters, and hedgehogs. Using rat anti-UCP-antibody, it is shown that UCP is located in the cristae of brown adipocytes mitochondria (UC-mitochondria) of all analyzed species, whereas mitochondria of nonadipose cells, i.e., C-mitochondria of endothelium, fibrocytes, smooth muscle cells, Schwann cells, axons of neural cells, and white blood cells, do not contain UCP. Cold stress in rats exposed to temperatures of +4 and -20 degrees C caused the amount of UCP per 1 micron 2 of mitochondria to more than double compared with room temperature-acclimated rats. This increase was especially dramatic on the outer mitochondrial membrane: fourfold more UCP molecules compared to the control rats. The ground cytoplasm of adipocytes showed very intensive labeling with RNase-gold complex, indicating that cytoplasm was an active site for protein synthesis, while the absence of UCP-labeling in ground cytoplasm was interpreted to mean that ground cytoplasm did not serve as a site for UCP synthesis. On the other hand, the protrusions of the outer mitochondrial membrane covered with ribosomes, clusters of UCP molecules, and clusters of RNase-gold particles supported the idea that UCP was one of the mitochondrial proteins synthesized on the ribosomes which were in contact with the outer mitochondrial membrane. After being synthesized there, UCP, which could be either extruded into intermembranous space or directed by lateral movement to intermembranous contact sites, was incorporated into inner mitochondrial membrane. Thus, UCP is imported using the so-called "cotranslational transport system."
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