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排序方式: 共有138条查询结果,搜索用时 15 毫秒
131.
Dennis V Pedersen Gregers R Andersen Dennis V Pedersen Lubka Roumenina Rasmus K Jensen Trine AF Gadeberg Chiara Marinozzi Capucine Picard Tania Rybkine Steffen Thiel Uffe BS Sørensen Cordula Stover Veronique Fremeaux-Bacchi Gregers R Andersen 《The EMBO journal》2017,36(8):1084-1099
Properdin (FP) is an essential positive regulator of the complement alternative pathway (AP) providing stabilization of the C3 and C5 convertases, but its oligomeric nature challenges structural analysis. We describe here a novel FP deficiency (E244K) caused by a single point mutation which results in a very low level of AP activity. Recombinant FP E244K is monomeric, fails to support bacteriolysis, and binds weakly to C3 products. We compare this to a monomeric unit excised from oligomeric FP, which is also dysfunctional in bacteriolysis but binds the AP proconvertase, C3 convertase, C3 products and partially stabilizes the convertase. The crystal structure of such a FP-convertase complex suggests that the major contact between FP and the AP convertase is mediated by a single FP thrombospondin repeat and a small region in C3b. Small angle X-ray scattering indicates that FP E244K is trapped in a compact conformation preventing its oligomerization. Our studies demonstrate an essential role of FP oligomerization in vivo while our monomers enable detailed structural insight paving the way for novel modulators of complement. 相似文献
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134.
Don R. Justesen Eleanor R. Adair Joseph C. Stevens Virginia Brucewolfe 《Bioelectromagnetics》1982,3(1):117-125
Three male and three female adults individually placed the ventral surface of the right and upright forearm against a 15-cm-diameter aperture in a wall of microwave-absorbent material. Tensecond exposures occurred to E-vector-vertically polarized, 2450-MHz-CW microwave (MW) fields. Comparable exposure to infrared (IR) waves was repeated with four of the six observers. Thresholds of detection of just-noticeable warming by MW and IR radiation were determined by the double-staircase psychophysical method. Although the exposed surface areas of male observers' arm were larger than those of female observers, thresholds of warming by either source of energy overlapped; the pooled means of irradiance at threshold are 26.7 mW/cm2 (MW) and 1.7 mW/cm2 (IR). Dosimetric measures on saline models indicated virtually perfect absorption of the incident IR, but nearly two-thirds of the MW energy was scattered. Accordingly, the 15-fold difference in means of MW and IR thresholds resolves to a 5-fold difference in threshold quantities of absorbed energy. In the light of the high correlation between thresholds of IR and MW irradiation (r = .97), it is concluded that the same set of superficial thermoreceptors was being stimulated, only less efficiently so, by the more deeply penetrating, more diffusely absorbed MW energy. 相似文献
135.
W H Stork J A Van Boxsel A F De Goeij AF+DEGOEIJ P L De Haseth PL+DEHASETH M Mandel 《Biophysical chemistry》1974,2(2):127-136
Binding of auramine O to poly-(methacrylic acid) (PMA) has been established over a large range of C0p/C0d values using spectroscopic methods (UV absorption and visible fluorescence emission spectra), equilibrium and sedimentation dialysis, potentiometric and viscosimetric titrations. All the results show qualitative agreement with those obtained previously with the system crystal violet-PMA although the binding seems to be less strong for auramine O than for crystal violet. From dialysis experiments binding isotherms were obtained at three different degrees of neutralization α'; at α' = 0.10 the results could be fitted to a Langmuir isotherm but at α' equal to 0.40 and 0.65 deviations with respect to such an isotherm occur. The results of potentiometric and viscosimetric titrations confirm that the conformational transition which the dye-free PMA exhibits upon ionization is affected by the dye binding. The region in which the conformational transion occurs is broadened and is less sharply defined in the presence of auramine O. 相似文献
136.
The interferon-induced enzyme 2-5A synthetase is shown to adenylate tRNA. Yeast tRNAPhe was incubated with the enzyme in the presence of double stranded RNA (in this case polyI-polyC) and ATP or deoxyATP. The reaction products were analyzed by ribonuclease T1 digestion of the tRNA, polyacrylamide gel electrophoresis and autoradiography. Using ATP, the 2-5A synthetase adds one, two or three AMP residues to the 3'-end of the tRNA whereas when dATP is replacing ATP, only one nucleotide unit is added. It is concluded that one of the mechanisms of the interferon-induced antiviral effect may be an inhibition of the translation process caused by an inactivation of tRNA molecules by a 2-5A synthetase catalyzed 2'-adenylation of the 3'-end. 相似文献
137.
The physiology of stringent factor (ATP:GTP 3'-diphosphotransferase) in Escherichia coli 总被引:4,自引:0,他引:4
The enzyme ATP:GTP 3'-diphosphotransferase catalyzes the transfer of the beta, gamma-pyrophosphate of ATP to the 3' position of GTP or GDP. The amounts of enzyme were measured in cell extracts of a relA+ strain of E. coli grown at different growth rates between 0.4 and 1.9 generations per hour, using precipitation with specific antibodies to purify the enzyme. The amount of enzyme was found to be a constant fraction of total protein at all growth rates corresponding to about 45 molecules of enzyme per genome equivalent of DNA. The purified enzyme has little catalytic activity by itself but has to be activated either by a complex of 70S ribosomes, mRNA and uncharged tRNA or by a solvent like ethanol at a concentration of about 20%. The kinetic constants of the enzyme for the transfer pyrophosphate from ATP to GTP in the ribosome-activated state were determined. The Vmax was estimated to be 140 mumol/min X mg at 37 degrees C and the S0.5 values for GTP and ATP were 0.35 and 0.53 mM, respectively. The reaction was estimated to have an equilibrium constant of about 300. In the pyrophosphate transfer from ATP to GDP the Vmax was estimated to be 90 mumol/min X mg at 37 degrees C and the S0.5 for GDP as 0.3 mM. During amino acid starvation of a relA+ strain of E. coli the amounts of enzyme and the catalytic capacity of the enzyme are sufficient to maintain the observed ppGpp levels in the cells at all growth rates. 相似文献
138.
Conni Lauritzen John Pedersen Mads T. Madsen Just Justesen Pia M. Martensen Søren W. Dahl 《Protein expression and purification》1998,14(3):434-442
An active form of rat dipeptidyl aminopeptidase I (DPPI, cathepsin C) was obtained by heterologous expression in insect cells. Baculoviruses carrying a cDNA sequence encoding the entire rat DPPI precursor was used to infect High Five cells in a serum-free medium. Recombinant DPPI (rDPPI) was secreted into the medium from which it was purified by a combination of ammonium sulfate fractionation, hydrophobic interaction chromatography (HIC), and ion-exchange chromatography. A polyhistidine-tagged form of the enzyme (HT-rDPPI) was purified from the medium by immobilized metal affinity chromatography (IMAC).In vivoactivation of native rat DPPI involves at least three chain cleavages per subunit and the ability of the expression system to imitate this processing was investigated. Both rDPPI and HT-rDPPI were secreted into the medium as unprocessed and inactive proenzymes and gradually converted into their active forms in the medium. This process was not completed at the time of harvest but mature enzyme processed similarly to native rat and human DPPI could be obtained by incubating the eluates from the HIC and IMAC columns at pH 4.5 and 5°C for 18–40 h. The yield of purified and matured enzyme was approximately 50 mg/liter, and it was shown that rDPPI and HT-rDPPI were active against both a dipeptide–p-nitroanilide substrate and human growth hormone N-terminally extended with an Ala-Glu dipeptide. 相似文献