An ATP-dependent system specific for degradation of long-lived proteins in permeabilized cells |
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| Authors: | D A Leonard H W Chen |
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| Institution: | Du Pont Experimental Station, Medical Products Department, E.I. du Pont de Nemours and Company, Wilmington, DE. |
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| Abstract: | We have characterized a digitonin-permeabilized cell system for the ATP-dependent degradation of endogenous long-lived proteins. Proteolysis requires Mg2+ and ATP hydrolysis. Other nucleotide triphosphates (CTP, UTP) can partially replace the ATP requirement. The enhanced rate of degradation of long-lived proteins in response to serum starvation is maintained in the permeabilized cell system and can be partially inhibited by lysosomal inhibitors. The maintenance of intracellular architecture and ease of manipulation of soluble components make the permeabilized cell system ideal for studying the proteolysis of both endogenous and exogenous substrates. |
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