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Connolly DT Witherbee BJ Melton MA Durley RC Grapperhaus ML McKinnis BR Vernier WF Babler MA Shieh JJ Smith ME Sikorski JA 《Biochemistry》2000,39(45):13870-13879
Chiral N,N-disubstituted trifluoro-3-amino-2-propanols represent a recently discovered class of compounds that inhibit the neutral lipid transfer activity of cholesteryl ester transfer protein (CETP). These compounds all contain a single chiral center that is essential for inhibitory activity. (R,S)SC-744, which is composed of a mixture of the two enantiomers, inhibits CETP-mediated transfer of [(3)H]cholesteryl ester ([(3)H]CE) from HDL donor particles to LDL acceptor particles with an IC(50) = 200 nM when assayed using a reconstituted system in buffer and with an IC(50) = 6 microM when assayed in plasma. Upon isolation of the enantiomers, it was found that the (R,+) enantiomer, SC-795, was about 10-fold more potent than the mixture, and that the (S,-) enantiomer, SC-794, did not have significant inhibitory activity (IC(50) > 0.8 microM). All of the activity of the (S,-)SC-794 enantiomer could be accounted for by contamination of this sample with a residual 2% of the highly potent (R,+) enantiomer, SC-795. The IC(50) of (R,+)SC-795, 20 nM, approached the concentration of CETP (8 nM) in the buffer assay. These chiral N,N-disubstituted trifluoro-3-amino-2-propanols were found to associate with both LDL and HDL, but did not disrupt overall lipoprotein structure. They did not affect the on or off rates of CETP binding to HDL disk particles. Inhibition was highly specific since the activities of phospholipid transfer protein and lecithin cholesterol acyl transferase were not affected. Competition experiments showed that the more potent enantiomer (R)SC-795 prevented cholesteryl ester binding to CETP, and direct binding experiments demonstrated that this inhibitor bound to CETP with high affinity and specificity. It is estimated, based on the relative concentrations of inhibitor and lipid in the transfer assay, that (R)SC-795 binds approximately 5000-fold more efficiently to CETP than the natural ligand, cholesteryl ester. We conclude that these chiral N,N-disubstituted trifluoro-3-amino-2-propanol compounds do not affect lipoprotein structure or CETP-lipoprotein recognition, but inhibit lipid transfer by binding to CETP reversibly and stereospecifically at a site that competes with neutral lipid binding. 相似文献
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This paper will review the challenges of developing a proteomics strategy. A key issue is the integration of the two-dimensional (2-D) gel platform with mass spectrometry measurements. The use of both matrix-assisted laser/desorption ionization (on off-line coupling) and electrospray (on-line) ionization are complementary. While the use of one-dimensional and 2-D gels are essential to many aspects of proteomics research (sample preparation, preliminary fractionation and quantitation, storage of protein components), the emergence of shotgun sequencing based on high performance liquid chromatography and tandem mass spectrometry offers a powerful new approach. The latter has particular utility in the characterization of low level samples and complex post-translational modifications. The development of capillary columns, such as 75 to 150 micron, that can be packed in a reproducible manner has been a key step in the development of high sensitivity liquid chromatography/mass spectrometry analysis. 相似文献
146.
For testing biological response of newborn Taiwan monkeys to infection of human adenovirus type 12 (Ad12) and simian virus 40 (SV40), 40 newborn Taiwan monkeys were inoculated with Ad12, Ad12, plus SV40, Ad12 supplemented with Ad12-induced hamster tumor tissue (Ad12 tumor) or control specimens (HeLa or African green monkey kidney cell lysate). Among them 26 survived including 8 newborn monkeys inoculated with control specimens. The survivors were observed for 4 years but no tumor was produced. The increase of body weight and intake of calories and protein in each test group during the first 12 wk were similar to those of corresponding control groups. Intrapulmonary inoculation of 108.2TCD50 of Ad12 with additional subcutaneous dose of Ad12 (108.8TCD50), Ad12 plus SV40 (108TCD50) or Ad12 plus Ad12 tumor killed 78% of newborn monkeys (7 of 9) in 18 days. The newborn could stand subcutaneous inoculation of SV40 (108TCD50) with 1 dose of Ad12 (108.8TCD50) or 3 doses of Ad12 (108.5, 108.5 and 108.2TCD50) at 24-hr intervals. When 108.8TCD50 or more Ad12 were inoculated, the virus could be isolated as late as 44 and 26 days from rectal and throat swab specimens respectively. The Ad12 neutralizing antibody in baby monkeys inoculated with multiple doses of Ad12 persisted, in low titer, longer than those injected with single high doses of Ad12, but anti-Ad12 T (tumor) antibody disappeared by 35 wk in both groups. Although SV40 antibody response was better than Ad12 antibody response in baby Taiwan monkeys, pre-infection of SV40 did not potentiate the production of anti-Ad12 T antibody. 相似文献
147.
H S Shieh N K Leimgruber M O Palmier T C Wun R M Leimgruber S S Abdel-Meguid 《Journal of molecular biology》1990,212(1):15-16
Crystals of a chymotrypsin inhibitor from Erythrina caffra seeds have been grown out of lithium sulfate, by the hanging drop method of vapor diffusion. The crystals belong to the rhombohedral space group R32, with a = 67.2 A and alpha = 99.4 degrees, and diffract to 3 A resolution. 相似文献
148.
H. S. Shieh 《Journal of fish biology》1978,12(1):13-18
Furunculosis was induced in brook trout, Salvelinus fontinalis , by experimental inoculation with Aeromonas salmonidda. The levels of blood enzymes were determined and the most significant modifications were observed for aldolase, creatine phosphokinase and ornithine carbamyl transferase. Alkaline phosphatase, glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase showed minor modification. There was no significant change of activity for acid phosphatase, cholinesterase, lactic dehydrogenase, phosphohexose iso-merase, isocitric dehydrogenase and aminopeptidase. 相似文献
149.
The intrinsic rate of nitrification was observed in a batch reactor by eliminating external and internal diffusional resistances. The former were minimized by means of intense agitation, and the latter by mechanical rupture of the activated sludge flocs using high mixer rotational speeds. The optimum temperature and pH for the intrinsic nitrification rate were found to be 30–35°C and 8.0, respectively. Initial ammonium concentration was found to have a strong effect on the value of the kinetic parameters of the Michaelis–Menten rate expression at low ammonium levels. However, at high initial concentrations both parameters attained a constant maximum value that is independent of the initial substrate level. 相似文献
150.