Calmodulin-NADH semidehydroascorbate oxidoreductase interactions of clathrin coated vesicles.
NADH semidehydroascorbate oxidoreductase activity is present in clathrin coated vesicles isolated from rat liver. The activity of the enzyme on the coated vesicles and Golgi apparatus but not that of endoplasmic reticulum is stimulated by calmodulin and is inhibited by three different drugs which are known inhibitors of calmodulin function including trifluoperizine, pimozide and R24571. Extraction of clathrin from the vesicles causes a decrease in activity which can be partially restored when the extracted clathrin is added back. Added calmodulin also restores much of the activity which is lost when the clathrin is removed and the specific activity of added pure calmodulin is similar to that of the crude clathrin on a protein basis. There is a decrease in enzyme activity if coated vesicles or Golgi apparatus are treated with a calcium antagonist (8-[N,N-diethylamino]-octyl-3,4,5-trimethoxybenzoate) (TMB-8). However, the enzyme activity can be recovered to that of the untreated control if calcium (6.0 mM) is added. An additive stimulatory effect on enzyme activity is also observed when both calcium (1.0 mM) and calmodulin (40 micrograms/ml) are present in the vesicles simultaneously. The results show that the NADH-semidehydroascorbate oxidoreductase of coated vesicles and Golgi apparatus have regulatory properties different from those of the microsomal electron transport system. Calmodulin-calcium control mediated through the semidehydroascorbate reductase, may be among the components that regulate Golgi apparatus secretion.
D000818:Animals; D002147:Calmodulin / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006056:Golgi Apparatus / Q000201:enzymology*; D007093:Imidazoles / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010868:Pimozide / Q000494:pharmacology; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology; D014268:Trifluoperazine / Q000494:pharmacology
D002147:Calmodulin; D002966:Clathrin; D004791:Enzyme Inhibitors; D007093:Imidazoles; D010868:Pimozide; D014268:Trifluoperazine; C031938:calmidazolium; D009247:NADH, NADPH Oxidoreductases; C023927:monodehydroascorbate reductase (NADH)