Interfering with the inhibitory mechanism of serpins: crystal structure of a complex formed between cleaved plasminogen activator inhibitor type 1 and a reactive-centre loop peptide

BACKGROUND: Plasminogen activator inhibitor type 1 (PAI-1) is an important endogenous regulator of the fibrinolytic system. Reduction of PAI-1 activity has been shown to enhance dissolution of blood clots. Like other serpins, PAI-1 binds covalently to a target serine protease, thereby irreversibly inactivating the enzyme. During this process the exposed reactive-centre loop of PAI-1 is believed to undergo a conformational change becoming inserted into beta sheet A of the serpin. Incubation with peptides from the reactive-centre loop transform serpins into a substrate for their target protease. It has been hypothesised that these peptides bind to beta sheet A, thereby hindering the conformational rearrangement leading to loop insertion and formation of the stable serpin-protease complex. RESULTS: We report here the 1.95 A X-ray crystal structure of a complex of a glycosylated mutant of PAI-1, PAI-1-ala335Glu, with two molecules of the inhibitory reactive-centre loop peptide N-Ac-TVASS-NH2. Both bound peptide molecules are located between beta strands 3A and 5A of the serpin. The binding kinetics of the peptide inhibitor to immobilised PAI-1-Ala335Glu, as monitored by surface plasmon resonance, is consistent with there being two different binding sites. CONCLUSIONS: This is the first reported crystal structure of a complex formed between a serpin and a serpin inhibitor. The localisation of the inhibitory peptide in the complex strongly supports the theory that molecules binding in the space between beta strands 3A and 5A of a serpin are able to prevent insertion of the reactive-centre loop into beta sheet A, thereby abolishing the ability of the serpin to irreversibly inactivate its target enzyme. The characterisation of the two binding sites for the peptide inhibitor provides a solid foundation for computer-aided design of novel, low molecular weight PAI-1 inhibitors.     

颗粒表面改性法制备PTC粉体的研究

本文介绍了一种通过液相颗粒表面改性技术添加二次掺杂物来制备ＰＴＣ粉体的新方法。即将烧结助熔剂Ａｌ２Ｏ３，ＳｉＯ２以及受主杂质ＭｎＯ采用溶液形式，包裹在ＢａＴｉＯ３基粉体颗粒表面进行改性，由此可以得到二次掺杂物Ａｌ２Ｏ３、ＳｉＯ２和ＭｎＯ分布均匀的ＰＴＣ粉体。     

Studies on purification and some properties of nisin from Lactococcus lactis subsp. lactis Al2

Protoporphyria is a genetic disorder in which patients may develop severe protoporphyrin-induced liver damage and require transplantation. Because unique problems occur in the perioperative period and because excess production of protoporphyrin by the bone marrow continues after liver transplantation, the efficacy of this procedure for protoporphyric liver disease is uncertain. We present follow-up of nine patients who underwent liver transplantation. Two patients died within 2 months of transplantation, one from complications of abdominal bleeding and the other from sepsis after bowel perforations. The remaining seven patients had follow-up at 14 months to 8 years after transplantation (mean, 3.8 years). Two of the seven had suffered skin burns from exposure to operating room lights, which healed without scarring. Three had axonal neuropathies in the postoperative period requiring prolonged mechanical ventilation, and motor defects persisted in two. Five patients had normal liver chemistries at follow-up (mean, 3.5 years), with liver biopsy results normal or showing mild portal triad abnormalities, but erythrocyte protoporphyrin levels remained significantly elevated (1,765 +/- 365 mcg/dL; normal, < 65). The other two patients, both of whom had rejection, cytomegalovirus infection, and biliary tract obstruction requiring endoscopic therapy, had a recurrence of protoporphyric liver disease as indicated by liver biopsy features. One died 5 years after transplantation from complications of the liver disease. The other was stable 3.3 years after transplantation and was being monitored for possible retransplantation. Thus, liver transplantation can be performed successfully in patients with protoporphyric liver disease, with intermediate survival rates comparable to the general transplant population. However, disease may recur in the graft, particularly if there are complications that cause cholestasis.     

漳泽水库最优兴利调度研究

漳泽水库供水能力下降。优化调度方式是提高水库效益的最有效途径。通过采用随机动态规划方法与模拟模型交互运算得到现行控制水位的水库优化调度图。采用同样方法对不同控制水位的水库运行方式进行优化调度，选出的规划水库最优控制水位为汛限水位９０４ ．５ ｍ ，正常高水位９０５ ．５ ｍ 。优化的控制水位方案在保障水库防洪安全的前提下可供给城市生活和工业用水６ ３００ 万ｍ３ ，较现行控制水位下的供水量提高１ ２５５ 万ｍ３ 。但农业用水仍然不足，应采取一定措施给予解决，如对灌区工程进行改造，因地制宜采取不同节水措施和污水回用等。     

电流变流体的研究进展

电流变流体是近年来逐渐受人们重视的一种功能材料，本文介绍其国内、外发展过程及研究现状。     

溶胶-凝胶法制备(Sr,Pb)TiO3纳米晶粉及表征

采用溶胶－凝胶法成功地合成了（Sr,Pb)TiO3纳米晶粉，并利用FTIR技术对凝胶的形成过程及350℃，1．5h煅烧获得的粉体材料进行了表征。实验以TG－DSC分析作为拟定煅烧温度的依据，以XRD作为煅烧产物的分析手段，最佳煅烧温度为600℃，所合成的（Sr,Pb)TiO3纳米晶粉的平均晶粒尺寸约为21nm，属钙钛矿型立方顺电相结构。     

利用分形几何确定多孔介质的孔尺寸分布

提出了利用分形几何确定多孔介质孔尺寸分布的方法。考虑多孔介质中孔间的屏蔽，采用此方法得到的孔尺寸分布范围比采用平行孔模型宽，分别采用本方法和平行孔模型测定γ－Al2O3催化剂的孔尺寸分布，构造简单立方体三维网络模型模拟压汞过程，并将模拟曲线与γ－Al2O3压汞实验曲线比较，结果表明，采用本方法所得孔尺雨分布优于平行孔模型所得孔尺寸分布。     

汽油机爆震强度的分析

分析了汽油机缸内气体爆震燃烧压力的频率特性并推导了爆震压力共振频率计算公式。研究表明：在爆震工况下，缸内压力在高频域中有几个共振峰，峰的大小表明了汽油机爆震强度。汽油机的表面振动在这些共振频率峰处也出现相应振动峰。表面振动峰与缸力压内峰值近似成正比。汽油机表面振动峰值可作为爆震强度的评价指标，该结论在ＣＡ１１０２汽油机上得到了证实。     

钨带灯的稳定性判据试验

我们通过对国产ＢＷ—１４００型真空钨带灯的稳定性判据进行研究，发现通过钨带灯的电流达到稳定的时间很短（小于５分钟），而钨带灯的灯电阻需要经过２０分钟后才能达到稳定，因此，用电流稳定性作为钨带灯的热平衡判据是不够的，更可靠的判据还应看灯电阻是否达到平衡。     

Temperature-regulated expression of the tac/lacl system for overproduction of a fungal xylanase in Escherichia coli

Temperature-regulated expression of recombinant proteins in the tac promoter (Ptac) system was investigated. Expression levels of fungal xylanase and cellulase from N. patriciarum in E. coli strains containing the natural lacI gene under the control of the Ptac markedly increased with increasing cultivation temperature in the absence of a chemical inducer. The specific activities (units per milligram protein of crude enzyme) of the fungal xylanase and cellulase produced from recombinant E. coli strain pop2136 grown at 42 degrees C were about 4.5 times higher than those of the cells grown at 23 degrees C and were even slightly higher when compared with cells grown in the presence of the inducer isopropyl beta-D-thiogalactopyranoside. The xylanase expression level in the temperature-regulated Ptac system was about 35% of total cellular protein. However, this system can not be applied to E. coli strains containing lacIq, which confers over production of the lac repressor, for high-level expression of recombinant proteins. In comparison with the lambda PL system, the Ptac-based xylanase plasmid in E. coli pop2136 gave a considerably higher specific activity of the xylanase than did the best lambda PL-based construct using the same thermal induction procedure. The high-level expression of the xylanase using the temperature-regulated Ptac system was also obtained in 10-litre fermentation studies using a fed-batch process. These results unambiguously demonstrated that the temperature-modulated Ptac system can be used for overproduction of some non-toxic recombinant proteins.