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Kinetics of product inhibition and mechanisms of lipoprotein lipase activation by apolipoprotein C-II 总被引:3,自引:0,他引:3
The kinetics of product inhibition of bovine milk lipoprotein lipase (LPL) were studied in a system of emulsified trioleoylglycerol (TG) at different fixed initial concentrations of oleic acid [( OA]0) without a fatty acid (FA) acceptor. In the absence of apolipoprotein C-II (C-II), the apparent Vmax and the nH(TG) (the slope of the corresponding Hill plot for TG) of 1.82 decreased by about 52% and [TG]0.5 increased 13-fold by raising the [OA]0 to 0.3 mM. At low [OA]0, product inhibition was competitive with respect to TG: the nH(OA) averaged 1.1, and [OA]0.5 was increased about 2-fold by TG. At the higher [OA]0, nH(OA) was 3.5, and TG had no effect on [OA]0.5. In the presence of 3 micrograms/mL C-II, the apparent Vmax was 4.3-7.1-fold higher than in its absence, and the nH(TG) was 2.45. Both parameters decreased by only 20-25%, and [TG]0.5 increased only 3-fold at an [OA]0 of 0.3 mM. Conversely, nH(OA) decreased by 35% and [OA]0.5 increased 6-fold by increasing TG concentrations. Similar kinetics were observed with very low density lipoproteins (VLDL). At saturating TG and varying C-II concentrations, nH(C-II) was 1.78, and product inhibition was found to be competitive with respect to C-II. At the [OA]0 employed, the FA had no effect on enzyme binding to TG emulsions, and there was no evidence that LPL catalyzes the reverse reaction. It is concluded that (a) the LPL kinetics are those of a multisite enzyme that probably has three high-affinity binding sites for TG, two for C-II, and four for OA.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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Rat pituitary tumors were induced by the monthly injection of high doses of estradiol valerate. Of 12 animals which received the estrogen, 10 developed tumors. From these 10 tumors, 4 cell lines were successfully established and they have been maintained in culture for over 18 months. Several clones have been isolated from these established cell lines. Three of the 4 cell lines produce tumors in females considerably faster than in males or castrated females. Tumor-bearing animals have significantly increased amounts of rat growth hormone and prolactin in their serum. The 4 established cell lines, as well as clones derived from them, and tumors obtained in situ by injection of these cells growing in culture or successive transplants have shown the presence of an estrogen-binding protein of high affinity and low capacity. This estrogen-binding protein is similar to that described in other target organs (uterus, mammary glands, etc). These cell lines will be used to study the mechanism of action of estrogen in target cells as well as the synthesis and secretion of pituitary hormones. 相似文献
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JC. Brisset F. Gazeau C. Corot N. Nighoghossian Y. Berthezène E. Canet-Soulas M. Wiart 《IRBM》2018,39(2):93-102