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排序方式: 共有816条查询结果,搜索用时 15 毫秒
41.
Taicatoxin, isolated from the venom of the Australian taipan snake Oxyuranus scutellatus, has been previously regarded as a specific blocker of high threshold Ca2+ channels in heart. Here we show that taicatoxin (in contrast to a range of other Ca2+ channel blockers) interacts with apamin-sensitive, small conductance, Ca2+-activated potassium channels on both chromaffin cells and in the brain. Taicatoxin displays high affinity recognition of 125I-apamin acceptor-binding sites, present on rat synaptosomal membranes (Ki = 1.45 +/- 0.22 nM) and also specifically blocks affinity-labeling of a 33-kDa 125I-apamin-binding polypeptide on rat brain membranes. Taicatoxin (50 nM) completely blocks apamin-sensitive after-hyperpolarizing slow tail K+ currents generated in rat chromaffin cells (mean block 97 +/- 3%, n = 12) while only partially reducing total voltage-dependent Ca2+ currents (mean block 12 +/- 4%, n = 6). In view of these findings, the use of taicatoxin as a specific ligand for Ca2+ channels should now be reconsidered. 相似文献
42.
The Rho family of small GTP-binding proteins are downregulated by an intrinsic GTPase, which is enhanced by GTPase-activating proteins (GAPs). RhoGAPs contain a single conserved arginine residue that has been proposed to be involved in catalysis. Here, the role of this arginine has been elucidated by mutagenesis followed by determination of catalytic and equilibrium binding constants using single-turnover kinetics, isothermal titration calorimetry, and scintillation proximity assays. The turnover numbers for wild-type, R282A, and R282K RhoGAPs were 5.4, 0.023, and 0.010 s-1, respectively. Thus, the function of this arginine could not be replaced by lysine or alanine. Nevertheless, the R282A mutation had a minimal effect on the binding affinity of RhoGAP for either Rho. GTP or Rho.GMPPNP, which confirms the importance of the arginine residue for catalysis as opposed to formation of the protein-protein complex. The R282A mutant RhoGAP still increased the hydrolysis rate of Rho.GTP by 160-fold, whereas the wild-type enzyme increased it by 38000-fold. We conclude that this arginine contributes half of the total reduction of activation energy of catalysis. In the presence of aluminum fluoride, the R282A mutant RhoGAP binds almost as well as the wild type to Rho.GDP, demonstrating that the conserved arginine is not required for this interaction. The affinity of wild-type RhoGAP for the triphosphate form of Rho is similar to that for Rho.GDP with aluminum fluoride. These last two observations show that this complex is not associated with the free energy changes expected for the transition state, although the Rho.GDP.AlF4-.RhoGAP complex might well be a close structural approximation. 相似文献
43.
Abstract Large additions of O2 during the pretreatment of modified or industrial kraft pulp with NO2 under conditions which favored an autocatalytic generation of NO2 from produced and added HNO3 led to a decreased viscosity after a subsequent oxygen bleaching, If the comparison was made at high kappa numbers. The difference was less apparent when the oxygen bleaching was extended. The yield of pulp at a given kappa number was reduced by large O2 additions. An Increased yield of nitric acid contributed to these effects. Hydrogen cyanide and ammonia were produced during the pretreatment in reactions between lignin and generated nitrogen oxides. Addition of O2 led to decreased yields of these products. 相似文献
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Purification and crystallization of complexes modeling the active state of the fragile histidine triad protein 总被引:1,自引:0,他引:1
Brenner C; Pace HC; Garrison PN; Robinson AK; Rosler A; Liu XH; Blackburn GM; Croce CM; Huebner K; Barnes LD 《Protein engineering, design & selection : PEDS》1997,10(12):1461-1463
Fragile histidine triad protein (Fhit) is a diadenosine triphosphate
(ApppA) hydrolase encoded at the human chromosome 3 fragile site which is
frequently disrupted in tumors. Reintroduction of FHIT coding sequences to
cancer cell lines with FHIT deletions suppressed the ability of these cell
lines to form tumors in nude mice even when the reintroduced FHIT gene had
been mutated to allow ApppA binding but not hydrolysis. Because this
suggested that the tumor suppressor activity of Fhit protein depends on
substrate-dependent signaling rather than ApppA catabolism, we prepared two
crystalline forms of Fhit protein that are expected to model its
biologically active, substrate-bound state. Wild-type and the His96Asn
forms of Fhit were overexpressed in Escherichia coli, purified to
homogeneity and crystallized in the presence and absence of ApppA and an
ApppA analog. Single crystals obtained by vapor diffusion against ammonium
sulfate diffracted X-rays to beyond 2.75 A resolution. High quality native
synchrotron X-ray data were collected for an orthorhombic and a hexagonal
crystal form.
相似文献
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A mixed general linear model analysis of the development of sleep-wake states was conducted on 37 high-risk preterm infants and replicated with a second cohort of 34 infants. Most dependent variables showed significant development over the preterm period: active sleep decreased, and active waking, quiet waking, and the organization of active sleep and quiet sleep increased over the preterm period in both cohorts. The amount of quiet sleep also increased over age, but this change was significant only for Cohort 1. Seven infant characteristics used as covariates had only minor effects. There were no significant differences in the developmental trajectories (slopes) of the two cohorts. The amounts of four variables differed between cohorts: Cohort 2 infants had less sleep-wake transition, more active sleep, less active sleep without REM, and more regular quiet sleep. These findings suggest that developmental patterns of sleep wake states are stable enough in the preterm period that deviant individual patterns might be used to identify infants with neurological problems. 相似文献
50.
HF Van Der Heijden WZ Zhan YS Prakash PN Dekhuijzen GC Sieck 《Canadian Metallurgical Quarterly》1998,85(2):525-529
The effects of the beta2-adrenoceptor agonist salbutamol (Slb) on isometric and isotonic contractile properties of the rat diaphragm muscle (Diamus) were examined. A loading dose of 25 microg/kg Slb was administered intracardially before Diamus excision to ensure adequate diffusion. Studies were then performed with 0.05 microM Slb in the in vitro tissue chamber. cAMP levels were determined by radioimmunoassay. Compared with controls (Ctl), cAMP levels were elevated after Slb treatment. In Slb-treated rats, isometric twitch and maximum tetanic force were increased by approximately 40 and approximately 20%, respectively. Maximum shortening velocity increased by approximately 15% after Slb treatment, and maximum power output increased by approximately 25%. During repeated isotonic activation, the rate of fatigue was faster in the Slb-treated Diamus, but both Slb-treated and Ctl Diamus fatigued to the same maximum power output. Still, endurance time during repetitive isotonic contractions was approximately 10% shorter in the Slb-treated Diamus. These results are consistent with the hypothesis that beta-adrenoceptor stimulation by Slb enhances Diamus contractility and that these effects of Slb are likely mediated, at least in part, by elevated cAMP. 相似文献