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1.
A simple approach to scan quickly a large protein sequence databasefor homology is described. The approach used is strictly dependenton the database organization. A database has been compiled inwhich protein sequences are grouped into families of closelyrelated proteins, each family being characterized by its averagedipeptide composition. A new entry in the database can be allocatedin a family by comparing its dipeptide composition with theaverage dipeptide composition of the families. 相似文献
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【目的】 比较嗜压和非嗜压微生物中蛋白质在氨基酸和二肽组成上的差异对嗜压蛋白稳定性机理的了解及在此基础上的定向改造具有重要意义。【方法】利用4种微生物全蛋白质组信息,统计了639对直系同源序列二级结构氨基酸组成及二肽组成并计算其偏差。【结果】结果表明:在β折叠和无规则卷曲中二者差异明显,β折叠中,嗜压蛋白含更多的缬氨酸,异亮氨酸,亮氨酸,更少的精氨酸,赖氨酸,天冬氨酸;无规则卷曲中,嗜压蛋白含更多的缬氨酸和异亮氨酸,更少的甘氨酸和脯氨酸。而嗜压蛋白存在更多的YM、MN、KD、QC、CI、MW、MM、CY、WQ、HC、RC和QH,更少TW、MS、VD、DH、YE、CT、MW、CF、CK、CM、MY、QI、TH、MQ、QQ和MC。【结论】二肽比氨基酸包含更多的结构和序列信息,更有利于了解嗜压蛋白稳定性机制及指导其定向改造。 相似文献
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基于最近邻居算法,从蛋白质一级序列出发,利用蛋白质序列氨基酸组成、二肤组成以及混合组成方法对蛋白质单聚体、二聚体、三聚体、四聚体、五聚体、六聚体和八聚体进行分类研究。结果表明:采用二肽组成编码方法的预洲效果最好,Jackknife检验和独立测试集检验的总体预测精度分别达到90.83%和95.48%,比相同数据集上基于伪氨基酸组成和组分耦合预测的方法提高了12和15个百分点;特别是对于五聚体蛋白,预测精度分别提高了90和50个百分点;说明二肽组成对于蛋白质四级结构分类研究是一种非常有效的特征提取方法。 相似文献
4.
The influence of polyhydric alcohols and carbohydrates on the thermostability, i.e., the heat inactivation kinetics, of Bacillus licheniformis alpha-amylase was studied in the temperature range 96 degrees to 130 degrees C. High concentrations (from 9 to 60 weight percent) of glycerol, sorbitol, mannitol, sucrose, or starch can markedly decrease the inactivation rate constant, k, and in the studied cases, this stabilizing effect grows stronger with increasing additive concentration. Statements about stabilization should, however, be specified carefully with respect to temperature, because E(A) is mostly altered likewise. For dissolved enzyme E(A) was almost always decreased in the presence of polyol or carbohydrate, whereas for immobilized enzyme it was augmented in each studied instance. The inactivation of dissolved enzyme can, in all the studied cases, be adequately described as a first-order process. Immobilized enzyme, however, shows biphasic then first-order inactivation kinetics, depending on the additive concentration and temperature. 相似文献
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Factors enhancing protein thermostability 总被引:22,自引:0,他引:22
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Relationship of protein flexibility to thermostability 总被引:11,自引:0,他引:11
M Vihinen 《Protein engineering》1987,1(6):477-480
Thermostability of proteins arises from the simultaneous effect of several forces, which in fact lead to decreased flexibility of the polypeptide chain. This is verified by flexibility indices, which are derived from normalized B-values of individual amino acids in several refined three-dimensional structures. Flexibility indices show that overall flexibility is reduced when thermostability is increased. Protein molecules require both flexibility and rigidity to function, but the higher the temperature optimum and stability the more rigid is the structure needed to compensate for increased thermal fluctuations. Flexibilities of proteins performing the same catalytic activity seem to be about the same at their temperature optima, but the more rigid thermostable proteins reach the flexibility of thermolabile proteins at higher temperatures. In several proteins such as allosteric enzymes, some local sites of flexibility are highly conserved. The relevance of reduced flexibility to overall stability of proteins is also discussed. Flexibility indices and profiles can be used in the design of more stable proteins by site-directed mutagenesis. 相似文献
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The composition of mengovirus virions produced by infected cells varies with the incubation temperature. Virons produced at 37.0 or 39.5 degrees contain four major polypeptides (alpha, beta, gamma, and delta) and one minor polypeptide (beta'). Virons produced at 31.5 degrees C contain two additional polypeptides (D1 and E). The virions of two temperature-sensitive (ts) and thermolabile mutants of mengovirus (ts25 and ts88) contain an increased amount of polypeptide beta', with a corresponding decrease in polypeptide beta when compared with the wild-type mengovirus. 相似文献
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Background
A large number of papers have been published on analysis of microarray data with particular emphasis on normalization of data, detection of differentially expressed genes, clustering of genes and regulatory network. On other hand there are only few studies on relation between expression level and composition of nucleotide/protein sequence, using expression data. There is a need to understand why particular genes/proteins express more in particular conditions. In this study, we analyze 3468 genes of Saccharomyces cerevisiae obtained from Holstege et al., (1998) to understand the relationship between expression level and amino acid composition. 相似文献10.
For the development of a method for the prediction of single point mutations substantially affecting protein thermostability, we studied the effect of the E85R and R82E mutations on the thermostability of thioredoxins from Escherichia coli (Trx) and Bacillus acidocaldarius (BacTrx), respectively. The basic method of investigation was the molecular dynamics simulation of 3D protein models in a particular solvent at different temperatures (300 and 373 K). Some thermolabile regions in Trx, BacTrx, and their mutants were revealed by analyzing the temperature effect on the molecular dynamics of the protein molecule. The effect of single point mutations on the temperature changes of the protein conformation mobility in several thermolabile regions was found. The results of the calculations are in accord with the experimental data indicating that the mutation E85R increases Trx thermostability, whereas the mutation R82E decreases BacTrx thermostability. The thermostability of these proteins was revealed to depend on ionic interactions between the thermolabile regions. The single point mutations change the parameters of these interactions and make them more favorable in the E85R-Trx mutant and less favorable in the R82E-BacTrx mutant. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 5; see also http: // www.maik.ru. 相似文献
11.
Seven hyper-stable multiple mutants have been constructed in staphylococcal nuclease by various combinations of eight different stabilizing single mutants. The stabilities of these multiple mutants determined by guanidine hydrochloride denaturation were 3.4 to 5.6 kcal/mol higher than that of the wild-type. Their thermal denaturation midpoint temperatures were 12.6 to 22.9 deg. C higher than that of the wild-type. These are among the greatest increases in protein stability and thermal denaturation midpoint temperature relative to the wild-type yet attained. There has been great interest in understanding how proteins found in thermophilic organisms are stabilized. One frequently cited theory is that the packing of hydrophobic side-chains is improved in the cores of proteins isolated from thermophiles when compared to proteins from mesophiles. The crystal structures of four single and five multiple stabilizing mutants of staphylococcal nuclease were solved to high resolution. No large overall structural change was found, with most changes localized around the sites of mutation. Rearrangements were observed in the packing of side-chains in the major hydrophobic core, although none of the mutations was in the core. It is surprising that detailed structural analysis showed that packing had improved, with the volume of the mutant protein's hydrophobic cores decreasing as protein stability increased. Further, the number of van der Waals interactions in the entire protein showed an experimentally significant increase correlated with increasing stability. These results indicate that optimization of packing follows as a natural consequence of increased protein thermostability and that good packing is not necessarily the proximate cause of high stability. Another popular theory is that thermostable proteins have more electrostatic and hydrogen bonding interactions and these are responsible for the high stabilities. The mutants here show that increased numbers of electrostatic and hydrogen bonding interactions are not obligatory for large increases in protein stability. 相似文献
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Corey A. Stevens Joanna Semrau Dragos Chiriac Morgan Litschko Robert L. Campbell David N. Langelaan Steven P. Smith Peter L. Davies John S. Allingham 《Protein science : a publication of the Protein Society》2017,26(10):1932-1941
Antifreeze proteins (AFPs) are a class of ice‐binding proteins that promote survival of a variety of cold‐adapted organisms by decreasing the freezing temperature of bodily fluids. A growing number of biomedical, agricultural, and commercial products, such as organs, foods, and industrial fluids, have benefited from the ability of AFPs to control ice crystal growth and prevent ice recrystallization at subzero temperatures. One limitation of AFP use in these latter contexts is their tendency to denature and irreversibly lose activity at the elevated temperatures of certain industrial processing or large‐scale AFP production. Using the small, thermolabile type III AFP as a model system, we demonstrate that AFP thermostability is dramatically enhanced via split intein‐mediated N‐ and C‐terminal end ligation. To engineer this circular protein, computational modeling and molecular dynamics simulations were applied to identify an extein sequence that would fill the 20‐Å gap separating the free ends of the AFP, yet impose little impact on the structure and entropic properties of its ice‐binding surface. The top candidate was then expressed in bacteria, and the circularized protein was isolated from the intein domains by ice‐affinity purification. This circularized AFP induced bipyramidal ice crystals during ice growth in the hysteresis gap and retained 40% of this activity even after incubation at 100°C for 30 min. NMR analysis implicated enhanced thermostability or refolding capacity of this protein compared to the noncyclized wild‐type AFP. These studies support protein backbone circularization as a means to expand the thermostability and practical applications of AFPs. 相似文献
13.
P Van Hoogevest A P Du Maine B De Kruijff J De Gier 《Biochimica et biophysica acta》1984,771(2):119-126
Glycophorin was incorporated into large unilamellar vesicles and the bilayer permeability was measured as a function of the lipid composition. In agreement with previous data (Van der Steen, A.T.M., De Kruijff, B. and De Gier, J. (1982) Biochim. Biophys. Acta 691, 13-23) it was found that glycophorin greatly increased the bilayer permeability of DOPC vesicles. This effect was observed for a large variety of phosphatidylcholines, differing in their fatty acid composition and homogeneity. In sharp contrast, it was observed that variations in the polar headgroups by incorporation of DOPE, DOPS and, to a lesser extent, cholesterol, into the DOPC/glycophorin vesicles restored the barrier function. These results are compared to the size of the particles, revealed by freeze-fracture electron microscopy on the glycophorin-containing bilayer and are discussed in the light of various types of lipid-protein interactions and protein aggregation state. 相似文献
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Given a novel protein it is very important to know if it is a DNA-binding protein, because DNA-binding proteins participate in the fundamental role to regulate gene expression. In this work, we propose a parallel fusion between a classifier trained using the features extracted from the gene ontology database and a classifier trained using the dipeptide composition of the protein. As classifiers the support vector machine (SVM) and the 1-nearest neighbour are used. Matthews's correlation coefficient obtained by our fusion method is approximately 0.97 when the jackknife cross-validation is used; this result outperforms the best performance obtained in the literature (0.924) using the same dataset where the SVM is trained using only the Chou's pseudo amino acid based features. In this work also the area under the ROC-curve (AUC) is reported and our results show that the fusion permits to obtain a very interesting 0.995 AUC. In particular we want to stress that our fusion obtains a 5% false negative with a 0% of false positive. Matthews's correlation coefficient obtained using the single best GO-number is only 0.7211 and hence it is not possible to use the gene ontology database as a simple lookup table. Finally, we test the complementarity of the two tested feature extraction methods using the Q-statistic. We obtain the very interesting result of 0.58, which means that the features extracted from the gene ontology database and the features extracted from the amino acid sequence are partially independent and that their parallel fusion should be studied more. 相似文献
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The influence of stream invertebrate composition at neighbouring sites on local assemblage composition 总被引:1,自引:0,他引:1
1. The composition of freshwater invertebrate assemblages at a location is determined by a range of physico‐chemical and biotic factors in the local environment, as well as larger‐scale spatial factors such as sources of recruits. We assessed the relative importance of the species composition of local neighbourhoods and proximal environmental factors on the composition of invertebrate assemblages. 2. Macroinvertebrate assemblages were sampled at 188 running‐water sites in the catchment of the River Rede, north‐east England. A total of 176 species were recorded. 3. Environmental data, in the form of 13 biotic and abiotic measurements that described stream physical structure, aquatic vegetation and water characteristics, were recorded for each site. Detrended correspondence analysis was then used to simplify nine of these stream environmental variables to create an index of stream structure. 4. The species composition of the invertebrate assemblages was related to the environmental variables, using an information theoretic approach. The impact of the species composition of neighbouring sites on each site was determined using Moran's I and autoregressive modelling techniques. 5. Species composition was primarily associated with water pH and stream structure. The importance of the species composition of neighbouring sites in determining local species assemblages differed markedly between taxa. The autoregressive component was low for Coleoptera, intermediate for Trichoptera and Plecoptera, and high for Ephemeroptera. 6. We hypothesise that the observed differences in the autoregressive component amongst these orders reflects variation in their dispersal abilities from neighbouring sites. 相似文献
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The bovine seminal plasma protein PDC-109 exerts an essential influence on the sperm cell plasma membrane during capacitation. However, by any mechanism, it has to be ensured that this function of the protein on sperm cells is not initiated too early, that is, upon ejaculation when PDC-109 and sperm cells come into first contact, but rather at later stages of sperm genesis in the female genital tract. To answer the question of whether changes of the bovine sperm lipid composition can modulate the effect of PDC-109 on sperm membranes, we have investigated the influence of PDC-109 on the integrity of (i) differently composed lipid vesicles and of (ii) membranes from human red blood cells and bovine spermatozoa. PDC-109 most effectively disturbed lipid membranes composed of choline-containing phospholipids and in the absence of cholesterol. The impact of the protein on lipid vesicles was attenuated in the presence of cholesterol or of noncholine-containing phospholipids, such as phosphatidylethanolamine or phosphatidylserine. An extraction of cholesterol from lipid or biological membranes using methyl-beta-cyclodextrin caused an increased membrane perturbation by PDC-109. Our results argue for a oppositional effect of PDC-109 during sperm cell genesis. We hypothesize that the lipid composition of ejaculated bull sperm cells allows a binding of PDC-109 without leading to an impairment of the plasma membrane. At later stages of sperm cell genesis upon release of cholesterol from sperm membranes, PDC-109 triggers a destabilization of the cells. 相似文献
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Polyacrylamide gel electrophoresis under reducing and non-reducing conditions, and immunoelectrophoretic analysis, revealed that several characteristic proteins disappear from the luminal fluid of the rat cauda epididymidis when it is maintained at body temperature. On SDS-PAGE gels prepared under reducing conditions, one Coomassie-blue staining band of Mr 18,000 disappeared and another of 52,000 was significantly reduced after only 6 days; bands of Mr 23,000, several in the Mr 34-38,000 range, one of Mr 48,000, and others of Mr 100-200,000 were eliminated or markedly reduced after 15 days at body temperature. Some were glycoprotein, as judged by their affinity for FITC-ConA. At 15 days after castration there was a broadly similar but rather more extensive disappearance of macromolecules, and of glycoproteins in particular, from caudal fluid. The fact that several similar proteins are diminished in or disappear from fluid or the cauda epididymidis maintained at body temperature, or after androgen withdrawal, raises the possibility that one or more such proteins play a role in sperm storage there. 相似文献