Inhibition of mutant troponin C activity by an intra-domain disulphide bond

Triggering of contraction in striated muscles involves a conformational transition in the N-terminal domain of troponin C, the calcium-binding component of thin filaments. We have designed a mutant troponin C in which the key conformational transition and the calcium-regulatory activity are reversibly blocked by the formation of a disulphide bridge. Our results may be applicable to other proteins of the same family of calcium-binding proteins.     

Isolation of an autotrophic ammonia-oxidizing marine archaeon

For years, microbiologists characterized the Archaea as obligate extremophiles that thrive in environments too harsh for other organisms. The limited physiological diversity among cultivated Archaea suggested that these organisms were metabolically constrained to a few environmental niches. For instance, all Crenarchaeota that are currently cultivated are sulphur-metabolizing thermophiles. However, landmark studies using cultivation-independent methods uncovered vast numbers of Crenarchaeota in cold oxic ocean waters. Subsequent molecular surveys demonstrated the ubiquity of these low-temperature Crenarchaeota in aquatic and terrestrial environments. The numerical dominance of marine Crenarchaeota--estimated at 10(28) cells in the world's oceans--suggests that they have a major role in global biogeochemical cycles. Indeed, isotopic analyses of marine crenarchaeal lipids suggest that these planktonic Archaea fix inorganic carbon. Here we report the isolation of a marine crenarchaeote that grows chemolithoautotrophically by aerobically oxidizing ammonia to nitrite--the first observation of nitrification in the Archaea. The autotrophic metabolism of this isolate, and its close phylogenetic relationship to environmental marine crenarchaeal sequences, suggests that nitrifying marine Crenarchaeota may be important to global carbon and nitrogen cycles.     

Site-directed mutagenesis reveals new and essential elements for iron-coordination of the sulfur oxygenase reductase from the acidothermophilic Acidianus tengchongensis

Previous study on refolding of sulfur oxygenase reductase (SOR) inclusion bodies from recombinant Escherichia coli showed that iron was critical to the activity of the SOR from Acidianus ambivalens. In this study, enzymatic assays showed that 2,2′-Dipyridyl, Tiron and 8-hydroxyquinoline, which are specific for chelating ferrous or ferric ions, strongly inhibited the activity of SOR from A. tengchongensis, suggesting that iron atom is essential for SOR activity. Alignment of several functionally identified S...     

Site-directed mutagenesis reveals new and essential elements for iron-coordination of the sulfur oxygenase reductase from the acidothermophilic Acidianus tengchongensis

Previous study on refolding of sulfur oxygenase reductase (SOR) inclusion bodies from recombinant Escherichia coli showed that iron was critical to the activity of the SOR from Acidianus ambivalens. In this study, enzymatic assays showed that 2,2′-Dipyridyl, Tiron and 8-hydroxyquinoline, which are specific for chelating ferrous or ferric ions, strongly inhibited the activity of SOR from A. tengchongensis, suggesting that iron atom is essential for SOR activity. Alignment of several functionally identified SORs and SOR-like sequences from genome database revealed a conserved, putative iron binding motif, H⁸⁶-X₃-H⁹⁰-X _n -E¹¹⁴-X _n -E¹²⁹ (numbering according to the Acidianus tengchongensis SOR sequence). Three mutants of SOR were generated by site-directed mutagenesis of H₈₆, H₉₀ and E₁₂₉ into phenylalanine or alanine residue in this study. Circular dichroism spectrum determination indicated that there was no change of the secondary structures of mutant SORs, H⁸⁶F, H⁹⁰F and E¹²⁹A, but all mutants were completely inactive. Through determination of iron contents we found that SOR mutants of H⁸⁶F, H⁹⁰F and E¹²⁹A completely or partially lost iron, while mutants of C³¹S, C¹⁰¹S, and C¹⁰⁴S (generated in a previous study) did not. This result indicated that H⁸⁶, H⁹⁰ and E¹²⁹ but not C³¹, C¹⁰¹, and C¹⁰⁴ were involved in binding to iron atom. Based on this and previous studies, it is proposed that the conserved motifs, C³¹-X _n -C¹⁰¹-X₂-C¹⁰⁴ and H⁸⁶-X₃-H⁹⁰-X₂₃-E¹¹⁴-X₁₄-(E/D)¹²⁹, are respectively for sulfur and molecular oxygen binding and activation. These two conserved motifs are essential elements for the SOR activity. Supported by National Natural Science Foundation of China (Grant Nos. 30670018, 30621005) and State Key Basic Research and Development Program of China (Grant No. 2004CB719602)     

一种新型碳材料——碳化物衍生碳的研究进展

评述了碳化物衍生碳的国内外研究现状,主要介绍了碳化物衍生碳的制备方法、结构与组成、在摩擦学及能源领域的应用,预测了碳化物衍生碳在今后的发展趋势。     

Evolution of carbides and carbon content in matrix of an ultra-high carbon sintered steel during heat treatment process

DTA, thermal expansion, XRD, and SEM were used to evaluate the effect of quenching temperature on the mechanical properties and microstructure of a novel sintered steel Fe-6Co-1Ni-5Cr-5Mo-1C. Lattice parameters and the mass fraction of carbon dissolved in the matrix of the steel quenched were investigated. It is discovered that the hardness of the steel increases with quenching temperature in the range of 840-900℃ and remains constant in the range of 900 to 1100℃. It decreases rapidly when the temperature is higher than 1100℃. The mass fraction of carbon dissolved in the matrix of the steel quenched at 840℃ is 0.38, but when the quenching temperature is increased to 1150℃, it increases to 0.98. The carbides formed during sintering are still present at grain boundaries and in the matrix of the steel quenched at low quenching temperatures, such as 840℃. When the quenching temperature is increased to 1150℃, most of the carbides at grain boundaries are dissolved with just a small amount of spherical M₂₃C₆ existing in the matrix of the quenched steel.     

一种新的电压高转换率的升压变流器

提出了一种新的具有高输入-输出电压转换率的升压变流器,该变流器含有两个升压电感和两个开关管,并采用一个耦合两个升压电感电流通道的辅助变压器,通过恒频控制可在较宽的负载范围和输入电压范围内实现对维持输出电压的调节。由一个1kW的实验原型电路证实了该变流器的性能。     

Substantial increase of protein stability by multiple disulphide bonds

Disulphide bonds can significantly stabilize the native structures of proteins. The effect is presumed to be due mainly to a decrease in the configurational chain entropy of the unfolded polypeptide. In phage T4 lysozyme, a disulphide-free enzyme, engineered disulphide mutants that crosslink residues 3-97, 9-164 and 21-142 are significantly more stable than the wild-type protein. To investigate the effect of multiple-disulphide bonds on protein stability, mutants were constructed in which two or three stabilizing disulphide bridges were combined in the same protein. Reversible thermal denaturation shows that the increase in melting temperature resulting from the individual disulphide bonds is approximately additive. The triple-disulphide variant unfolds at a temperature 23.4 degrees C higher than wild-type lysozyme. The results demonstrate that a combination of disulphide bonds, each of which contributes to stability, can achieve substantial overall improvement in the stability of a protein.     

酶法制备明胶的新工艺研究

阐述了一种酶法制备明胶的新工艺，并对酶用量、酶解温度、酶解时间、溶胶温度、溶胶初始pH值等参数进行了优化。同时，还测定了产品的理化性质。研究表明新工艺生产周期短(7～10d)，环境污染小，产率高。     

二氧化碳向碳酸二甲酯的选择催化转化

报道了从CO2和甲醇为原料直接合成碳酸二甲酯的研究进展。表明使用有机金属催化剂、耦合反应过程、超临界反应条件、产物的原位分离是实现CO2向碳酸二甲酯选择催化转化的重要方向。     

硅院转制:寻找新"奶酪"

2001年5月9日,由山东省硅酸盐研究设计院联合香港利通国际有限公司、中国国际热能工程公司等5家法人及22名自然人共同发起设立的山东硅苑新材料科技股份有限公司在淄博高新区宣告成立,有着43年历史的山东省硅酸盐研究设计院由此脱下“官帽”,成为我省97家省属科研院所整体改制第一家。“一个企业和一个人一样,生命到一定时候就会僵化,如果此时给它换换肌体、换换血液,就能量新延长它的生命,焕发新的活力。”山东硅苑新材料科技股份有限公司董事长、总经理殷书建这样诠释引人注目的硅院转制。     

在经典光学统计下频率上转换过程中相对平均光子数的演变

讨论了在用相干光和非相干光入射情况下，频率上转换过程中相对平均光子数随时间或空间的变化以及它们之间的差别。     

Structures of human insulin-degrading enzyme reveal a new substrate recognition mechanism

Insulin-degrading enzyme (IDE), a Zn2+-metalloprotease, is involved in the clearance of insulin and amyloid-beta (refs 1-3). Loss-of-function mutations of IDE in rodents cause glucose intolerance and cerebral accumulation of amyloid-beta, whereas enhanced IDE activity effectively reduces brain amyloid-beta (refs 4-7). Here we report structures of human IDE in complex with four substrates (insulin B chain, amyloid-beta peptide (1-40), amylin and glucagon). The amino- and carboxy-terminal domains of IDE (IDE-N and IDE-C, respectively) form an enclosed cage just large enough to encapsulate insulin. Extensive contacts between IDE-N and IDE-C keep the degradation chamber of IDE inaccessible to substrates. Repositioning of the IDE domains enables substrate access to the catalytic cavity. IDE uses size and charge distribution of the substrate-binding cavity selectively to entrap structurally diverse polypeptides. The enclosed substrate undergoes conformational changes to form beta-sheets with two discrete regions of IDE for its degradation. Consistent with this model, mutations disrupting the contacts between IDE-N and IDE-C increase IDE catalytic activity 40-fold. The molecular basis for substrate recognition and allosteric regulation of IDE could aid in designing IDE-based therapies to control cerebral amyloid-beta and blood sugar concentrations.