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1.
Substantial increase of protein stability by multiple disulphide bonds 总被引:25,自引:0,他引:25
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. 相似文献
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Defective co-translational formation of disulphide bonds in protein disulphide-isomerase-deficient microsomes 总被引:35,自引:0,他引:35
The formation of disulphide bonds in mammalian secretory and cell-surface proteins occurs in the lumen of the endoplasmic reticulum and is believed to be catalysed by the enzyme protein disulphide-isomerase (PDI). The evidence for this physiological role for PDI is circumstantial and relates to the cell and tissue distribution of the enzyme, its developmental behaviour and its catalytic properties in vitro. A clear requirement for PDI in the correct folding or assembly of disulphide-bonded proteins during biosynthesis has not been demonstrated. We have prepared dog pancreas microsomes which are deficient in soluble lumenal proteins, including PDI, but which are still able to translocate and process proteins synthesized in vitro. Using the formation of intramolecular disulphide bonds during the in vitro synthesis of gamma-gliadin, a wheat storage protein, as a model, we have demonstrated that these microsomes are defective in co-translational formation of disulphide bonds. Reconstitution of these microsomes with purified PDI reverses this defect. 相似文献
3.
Role of ATP and disulphide bonds during protein folding in the endoplasmic reticulum. 总被引:24,自引:0,他引:24
Being topologically equivalent to the extracellular space, the lumen of the endoplasmic reticulum (ER) provides a unique folding environment for newly synthesized proteins. Unlike other compartments in the cell where folding occurs, the ER is oxidizing and therefore can promote the formation of disulphide bonds. The reducing agent dithiothreitol, when added to living cells, inhibits disulphide formation with profound effects on folding. Taking advantage of this effect, we demonstrate here that folding of influenza haemagglutinin is energy dependent. Metabolic energy is required to support the correct folding and disulphide bond formation in this well characterized viral glycoprotein, to rescue misfolded proteins from disulphide-linked aggregates, and to maintain the oxidized protein in its folded and oligomerization-competent state. 相似文献
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Transport of the essential nutrient phosphorus--primarily in the form of orthophosphate--into cells and organelles is highly specific. This is exemplified by the uptake of phosphate or its close analogue arsenate by bacterial cells by way of a high affinity active transport system dependent on a phosphate-binding protein; this system is unable to recognize other inorganic oxyanions and is, moreover, distinct from the one for sulphate transport. The phosphate-binding protein is a member of a family of periplasmic proteins acting as initial high-affinity receptors for the osmotic shock-sensitive active transport systems or permeases for various sugars, amino acids, oligopeptides, and oxyanions. We report here the highly refined 1.7 A resolution X-ray structure of the liganded form of the phosphate-binding protein. The structure reveals the atomic features responsible for phosphate selectivity, either in monobasic or dibasic form, and the exclusion of sulphate. These features are fundamental to understanding phosphate transport systems and molecular recognition of charged substrates or ions in other biological processes. 相似文献
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Sequence of protein disulphide isomerase and implications of its relationship to thioredoxin 总被引:17,自引:0,他引:17
The formation of disulphide bonds is essential to the structure and function of proteins. These bonds rapidly form either cotranslationally or immediately post-translationally in the lumen of the endoplasmic reticulum. Native disulphide pairing for such proteins has been achieved in vitro; however, the rates of reassembly are slow and the conditions non-physiological. To account for these observations, Anfinsen et al. proposed that a 'disulphide interchange protein' was the in vivo catalyst of disulphide bond rearrangement. Other groups discovered an activity with similar characteristics that catalysed the reductive cleavage of insulin and may be associated with insulin degradation, although this result has been disputed. The enzyme involved, protein disulphide isomerase (PDI; EC 5.3.4.1), may be the in vivo catalyst of disulphide bond formation. Here we describe the sequence of cloned rat liver PDI complementary DNA which predicts a protein with two distinct regions homologous with Escherichia coli thioredoxin, a known cofactor in oxidation-reduction reactions. Each of these regions contains the presumed active site sequence Trp-Cys-Gly-His-Cys-Lys, suggesting that PDI, similar in action to thioredoxin, catalyses disulphide bond interchange via an internal disulphide-sulphydryl interchange. The cDNA predicts a signal peptide consistent with the view that PDI is a luminal endoplasmic reticulum protein. PDI messenger RNA, although ubiquitous, is more highly concentrated in secretory cells. 相似文献
8.
A Alon I Grossman Y Gat VK Kodali F DiMaio T Mehlman G Haran D Baker C Thorpe D Fass 《Nature》2012,488(7411):414-418
Protein stability, assembly, localization and regulation often depend on the formation of disulphide crosslinks between cysteine side chains. Enzymes known as sulphydryl oxidases catalyse de novo disulphide formation and initiate intra- and intermolecular dithiol/disulphide relays to deliver the disulphides to substrate proteins. Quiescin sulphydryl oxidase (QSOX) is a unique, multi-domain disulphide catalyst that is localized primarily to the Golgi apparatus and secreted fluids and has attracted attention owing to its overproduction in tumours. In addition to its physiological importance, QSOX is a mechanistically intriguing enzyme, encompassing functions typically carried out by a series of proteins in other disulphide-formation pathways. How disulphides are relayed through the multiple redox-active sites of QSOX and whether there is a functional benefit to concatenating these sites on a single polypeptide are open questions. Here we present the first crystal structure of an intact QSOX enzyme, derived from a trypanosome parasite. Notably, sequential sites in the disulphide relay were found more than 40?? apart in this structure, too far for direct disulphide transfer. To resolve this puzzle, we trapped and crystallized an intermediate in the disulphide hand-off, which showed a 165° domain rotation relative to the original structure, bringing the two active sites within disulphide-bonding distance. The comparable structure of a mammalian QSOX enzyme, also presented here, shows further biochemical features that facilitate disulphide transfer in metazoan orthologues. Finally, we quantified the contribution of concatenation to QSOX activity, providing general lessons for the understanding of multi-domain enzymes and the design of new catalytic relays. 相似文献
9.
The conformation of membranes 总被引:18,自引:0,他引:18
R Lipowsky 《Nature》1991,349(6309):475-481
Membranes composed of amphiphilic molecules are highly flexible surfaces that determine the architecture of biological systems and provide a basic structural element for complex fluids such as microemulsions. Physical theories have been developed to describe many aspects of their conformational behaviour, such as the preferred shapes and shape transformations of closed vesicles, and the shape fluctuations, random-surface configurations, and adhesion and unbinding of interacting membranes. Understanding of these phenomena has been much improved through fruitful interactions between theory and experiment. 相似文献
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利用集团模型探讨了硫对砷化镓(110)理想表面悬挂键的饱和作用,着重于定性讨论硫去除禁带内砷化镓表面态的可能性,这有助于了解硫钝化砷化镓表面的机理,计算采用电荷自洽的EHT方法,结果表明,硫确能与表面砷化镓原子形成较强的共价键,并把悬挂键形成的表面态移出禁带,其中硫与镓的相互作用更为强烈。 相似文献
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Sulphate sequestered in the sulphate-binding protein of Salmonella typhimurium is bound solely by hydrogen bonds 总被引:2,自引:0,他引:2
An important question in understanding substrate binding by proteins is how charged groups are stabilized in the absence of their solvation shell. We have addressed this question here by solving the structure of the sulphate-binding protein of Salmonella typhimurium with bound substrate at 2.0 A resolution. The results are remarkable in that the charged oxygen atoms of the sulphate molecule, which is buried and completely inaccessible to the solvent, are not stabilized by the formation of salt-bridges but by hydrogen bonds donated by specific residues of the protein. These hydrogen bonds are in turn coupled via peptide units to several resonating hydrogen bonding systems. These findings may be of general significance for the role of electrostatic interactions in protein structure and function. 相似文献
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研究模糊神经网络在可转换债券价格预测中的应用;并对万科转债(125002)进行价格预测,通过实际数据检验确定该方法的可行性;从而为可转换债券价格预测提供了一种行之有效的方法。 相似文献
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采用熔盐法合成了ZnTiO3粉体,研究了熔盐体系、氧化物掺杂、及煅烧制度对产物相结构的影响.结果表明:以NaCl/KCl复合熔盐可合成近于纯相的六方ZnTiO3.MgO、V2O5和W2O3掺杂获得的粉体主晶相均为ZnTiO3;MgO掺杂量为0.3时相稳定效果最好;而V2O5和W2O3掺杂量的变化对相结构影响不明显.在800~850℃获得了稳定的六方ZnTiO3相,保温时间的延长会促进ZnTiO3的分解. 相似文献
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特殊功能材料是一些具有优良电学、磁学、学、热学、学、力学、化学、物医学功能,在各类高科技领域到泛应用.正电子湮没技术是一种对材料微结构特别有效探测技术,特别是对各种缺陷、空位和微孔尤为灵敏,通过正电子湮没寿命谱、多普勒展宽谱和慢正电子束技术,通过分析正电子湮没参数可以获材料从表面到内部缺陷分布信息和随外部物理和化学条件变化、引起微结构变化.本文选取几种特殊材料正电子湮没实验结果来分析材料内部微结构,表明正电子湮没谱学是一种独特研究微观结构方法. 相似文献
16.
Wen Xiang-e Li Shi-qing Ma Li Yan He-ping Wang Zhu Wang Shao-jie Guo Cheng-zhan Zhao Hai-ge 《武汉大学学报:自然科学英文版》1999,4(3):290-294
Two samples of high purity InP extracted from the same wafer were examined by positron annihilation spectrum analysis after
having been, processed by means of thermal Neutron Transmutation Doping (NTD). Compared with the as grown sample with an average
positron lifetime of 246 ps at 300 K, the high dose doped one has an average lifetime of 251 ps and the lower dose doped one
248 ps measured under the same condition, indicating that some defects have been introduced in the NTD process. Annealing
experimental results show a steady decrease in the average lifetime with increasing annealing temperature up to 550°C. And
a peak in lifetime curve around 500°C was observed which may be attributed to defects related structure conversion. Temperature
experiments conducted on the low dose doped sample from 150K to 290 K suggest the existence of vacancy-impurity complex which
have given rise to an abnormal reduction of average lifetime with increasing temperature. Also a n-type InP sample (A61) was
irradiated with thermal neutrons in another reactor and the lifetime results display an increase of 15 ps. Furthermore, to
study epithermal neutron irradiation effects on InP, measurements were performed on an n-type InP sample (N119) along with
one p-type sample (P118) after having been irradiated with high fluence of epithermal neutrons. The former has an average
lifetime of 262 ps and the latter 247 ps after irradiation. The results prove that on some occassion epithermal neutrons can
produce sizable defects in InP.
Foundation item: Supported by the Science Foundation of Hubei Province (203980532)
Biography: WEN Xiang-e (1976-), male, Master candidate, Research direction; majar research interest is defects in semiconductor
materials using positron annihilation spectroscopy. 相似文献
17.
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. 相似文献
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《佛山科学技术学院学报(自然科学版)》2015,(6)
对我国国债市场上的跨市场交易品种的跨市套利交易机会进行研究,结果表明,在回购交易市场上,回购利率的差异在大多数情况下可以给作为交易所市场会员的证券公司投资者带来无风险套利利润,并且不会带来较大的流动性风险,但是,若要使套利利润达到一定量(比如10 000元),则所需的套利交易量将大大增大,从而使得套利交易面临较大的流动性风险。而对于非交易所市场会员的其他投资者来说,由于交易需要支付大量的佣金,而使得套利交易的机会大大减少。 相似文献
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Random coil conformation of polypeptide hormone precursor protein in endocrine cells 总被引:2,自引:0,他引:2
A G Pearse 《Nature》1969,221(5187):1210-1211
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