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161.
Environment-specific amino acid substitution tables: tertiary templates and prediction of protein folds. 总被引:4,自引:0,他引:4
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J Overington D Donnelly M S Johnson A Sali T L Blundell 《Protein science : a publication of the Protein Society》1992,1(2):216-226
The local environment of an amino acid in a folded protein determines the acceptability of mutations at that position. In order to characterize and quantify these structural constraints, we have made a comparative analysis of families of homologous proteins. Residues in each structure are classified according to amino acid type, secondary structure, accessibility of the side chain, and existence of hydrogen bonds from the side chains. Analysis of the pattern of observed substitutions as a function of local environment shows that there are distinct patterns, especially for buried polar residues. The substitution data tables are available on diskette with Protein Science. Given the fold of a protein, one is able to predict sequences compatible with the fold (profiles or templates) and potentially to discriminate between a correctly folded and misfolded protein. Conversely, analysis of residue variation across a family of aligned sequences in terms of substitution profiles can allow prediction of secondary structure or tertiary environment. 相似文献
162.
Crystal structure of the Y52F/Y73F double mutant of phospholipase A2: increased hydrophobic interactions of the phenyl groups compensate for the disrupted hydrogen bonds of the tyrosines. 总被引:3,自引:1,他引:2
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C. Sekharudu B. Ramakrishnan B. Huang R. T. Jiang C. M. Dupureur M. D. Tsai M. Sundaralingam 《Protein science : a publication of the Protein Society》1992,1(12):1585-1594
The enzyme phospholipase A2 (PLA2) catalyzes the hydrolysis of the sn-2 ester bond of membrane phospholipids. The highly conserved Tyr residues 52 and 73 in the enzyme form hydrogen bonds to the carboxylate group of the catalytic Asp-99. These hydrogen bonds were initially regarded as essential for the interfacial recognition and the stability of the overall catalytic network. The elimination of the hydrogen bonds involving the phenolic hydroxyl groups of the Tyr-52 and -73 by changing them to Phe lowered the stability but did not significantly affect the catalytic activity of the enzyme. The X-ray crystal structure of the double mutant Y52F/Y73F has been determined at 1.93 A resolution to study the effect of the mutation on the structure. The crystals are trigonal, space group P3(1)21, with cell parameters a = b = 46.3 A and c = 102.95 A. Intensity data were collected on a Siemens area detector, 8,024 reflections were unique with an R(sym) of 4.5% out of a total of 27,203. The structure was refined using all the unique reflections by XPLOR to a final R-factor of 18.6% for 955 protein atoms, 91 water molecules, and 1 calcium ion. The root mean square deviation for the alpha-carbon atoms between the double mutant and wild type was 0.56 A. The crystal structure revealed that four hydrogen bonds were lost in the catalytic network; three involving the tyrosines and one involving Pro-68. However, the hydrogen bonds of the catalytic triad, His-48, Asp-99, and the catalytic water, are retained. There is no additional solvent molecule at the active site to replace the missing hydroxyl groups; instead, the replacement of the phenolic OH groups by H atoms draws the Phe residues closer to the neighboring residues compared to wild type; Phe-52 moves toward His-48 and Asp-99 of the catalytic diad, and Phe-73 moves toward Met-8, both by about 0.5 A. The closing of the voids left by the OH groups increases the hydrophobic interactions compensating for the lost hydrogen bonds. The conservation of the triad hydrogen bonds and the stabilization of the active site by the increased hydrophobic interactions could explain why the double mutant has activity similar to wild type. The results indicate that the aspartyl carboxylate group of the catalytic triad can function alone without additional support from the hydrogen bonds of the two Tyr residues. 相似文献
163.
K. Harata M. Muraki Y. Hayashi Y. Jigami 《Protein science : a publication of the Protein Society》1992,1(11):1447-1453
The three-dimensional structure of a modified human lysozyme (HL), Glu 53 HL, in which Asp 53 was replaced by Glu, has been determined at 1.77 A resolution by X-ray analysis. The backbone structure of Glu 53 HL is essentially the same as the structure of wild-type HL. The root mean square difference for the superposition of equivalent C alpha atoms is 0.141 A. Except for the Glu 53 residue, the structure of the active site region is largely conserved between Glu 53 HL and wild-type HL. However, the hydrogen bond network differs because of the small shift or rotation of side chain groups. The carboxyl group of Glu 53 points to the carboxyl group of Glu 35 with a distance of 4.7 A between the nearest carboxyl oxygen atoms. A water molecule links these carboxyl groups by a hydrogen bond bridge. The active site structure explains well the fact that the binding ability for substrates does not significantly differ between Glu 53 HL and wild-type HL. On the other hand, the positional and orientational change of the carboxyl group of the residue 53 caused by the mutation is considered to be responsible for the low catalytic activity (ca. 1%) of Glu 53 HL. The requirement of precise positioning for the carboxyl group suggests the possibility that the Glu 53 residue contributes more than a simple electrostatic stabilization of the intermediate in the catalysis reaction. 相似文献
164.
X-ray crystal structures of the oxidized and reduced forms of the rubredoxin from the marine hyperthermophilic archaebacterium Pyrococcus furiosus. 总被引:10,自引:8,他引:2
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M. W. Day B. T. Hsu L. Joshua-Tor J. B. Park Z. H. Zhou M. W. Adams D. C. Rees 《Protein science : a publication of the Protein Society》1992,1(11):1494-1507
The structures of the oxidized and reduced forms of the rubredoxin from the archaebacterium, Pyrococcus furiosus, an organism that grows optimally at 100 degrees C, have been determined by X-ray crystallography to a resolution of 1.8 A. Crystals of this rubredoxin grow in space group P2(1)2(1)2(1) with room temperature cell dimensions a = 34.6 A, b = 35.5 A, and c = 44.4 A. Initial phases were determined by the method of molecular replacement using the oxidized form of the rubredoxin from the mesophilic eubacterium, Clostridium pasteurianum, as a starting model. The oxidized and reduced models of P. furiosus rubredoxin each contain 414 nonhydrogen protein atoms comprising 53 residues. The model of the oxidized form contains 61 solvent H2O oxygen atoms and has been refined with X-PLOR and TNT to a final R = 0.178 with root mean square (rms) deviations from ideality in bond distances and bond angles of 0.014 A and 2.06 degrees, respectively. The model of the reduced form contains 37 solvent H2O oxygen atoms and has been refined to R = 0.193 with rms deviations from ideality in bond lengths of 0.012 A and in bond angles of 1.95 degrees. The overall structure of P. furiosus rubredoxin is similar to the structures of mesophilic rubredoxins, with the exception of a more extensive hydrogen-bonding network in the beta-sheet region and multiple electrostatic interactions (salt bridge, hydrogen bonds) of the Glu 14 side chain with groups on three other residues (the amino-terminal nitrogen of Ala 1; the indole nitrogen of Trp 3; and the amide nitrogen group of Phe 29). The influence of these and other features upon the thermostability of the P. furiosus protein is discussed. 相似文献
165.
黄淮海平原高产田作物群体结构特征 总被引:9,自引:5,他引:4
本文利用实测资料分析了高产田作物群体结构特征。精播高产栽培麦田春季最大蘖数12×10~6/ha,群体最大叶面积指数5.5—6.0,成穗4.5—5.25×10~6/ha。传统高产栽培麦田春季最大蘖数15—18×10~6/ha,群体最大叶面积指数6.0—6.5,成穗6.0—7.5×10~6/ha。小麦营养生长与生殖生长期叶日积比,精播高产栽培麦田是1∶0.89,传统高产栽培麦田为1∶0.73。夏玉米种植密度主要受叶倾角的影响。紧凑型玉米品种的叶倾角大于65°,种植密度7.5—8.25×10~4/ha;平展型玉米品种的叶倾角小于50°,种植密度5.25—6.0×10~4/ha。 相似文献
166.
Primary structure of -chain of pigeon is presented. It was determined by amino acid sequence analysis of intact -chain and its peptides obtained by the enzymatic and chemical cleavage. Comparison of amino acid sequence of the chain with other available data shows 14 Ile, 61 Lys, and 113 Ile as residues specific to pigeon. One important replacement at 11 contact is 55 MetSer. 相似文献
167.
Igor A. Krasheninnikov Anton A. Komar Ivan A. Adzhubei 《Journal of Protein Chemistry》1991,10(5):445-453
Examination of nascent globin peptides accumulatingin vitro during globin synthesis in rabbit reticulocyte lysates was carried out. A view was supported that nonrandom distribution of codons with different usage frequencies in mRNA may determine the messenger's translation kinetics. Regions of reduced translation of - and -globin polypeptide chains were localized, and the cotranslational protein-folding model suggested previously was substantiated. An active conjunction of synthesis and folding of proteins was proposed as one of the main destinations of a translation nonuniformity. 相似文献
168.
On artificial polyethylene membranes providing a thigmotropic signal, uredospores of the broad bean rust fungus Uromyces viciae-fabae differentiated a series of infection structures which in nature are necessary to invade the host tissue through the stomata. Within 24 h germ tubes, appressoria, substomatal vesicles, infection hyphae and haustorial mother cells were developed successively. Alterations in protein metabolism during infection structure differentiation of this obligate plant pathogen were analyzed in the absence of the host plant by high resolution two-dimensional polyacrylamide gel electrophoresis (2-DE) and silver staining. The norm pattern representing the 2-DE protein patterns of the whole developmental sequence of infection structures of U. viciae-fabae showed 733 spots. During infection structure differentiation 55 proteins were newly formed, altered in quantity, or disappeared. Major alterations in the protein pattern occurred during uredospore germination and when infection hyphae were formed. Uredospore germination was characterized by a decrease of acidic proteins and an increase mainly of proteins with isoelectric points ranging from weakly acidic to basic.Abbreviations 2-DE
two-dimensional polyacrylamide gel electrophoresis
- DAPI
4,6-diamino-phenylindol
- kDa
kilo Dalton
- pl
isoelectric point
- PMSF
phenylmethylsulfonyl fluoride
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis 相似文献
169.
Species number, species abundance and body length of arboreal arthropods associated with an Australian rainforest tree 总被引:1,自引:0,他引:1
Abstract.
- 1 The species number, the abundance per species and the body length of arthropods foraging within the crowns of an over-storey rainforest tree from Australia, Argyrodendron actinophyllum (Sterculiaceae), were investigated by interception trap sampling and restricted canopy fogging. Emphasis was placed upon the interpretation of trap data. Arthropods were trapped continuously day and night, over a 2-year period and the final analyses examined the attributes of 759 species which represented 20,500 individuals.
- 2 The proportion of‘rare’species (Le. collected once) intercepted was high (35.7%), although lower than in other similar rainforest surveys. Neither the α log-series nor the log-normal distribution could be fitted to the relationship between number of species and number of individuals, since the number of rare species was much higher than predicted and the mode of the distribution could not be identified. The proportion of rare species was higher in fogging collections (452%) than in trap collections.
- 3 The data are compared with a study of Bornean arboreal beetles, obtained by fogging trees during a single sampling event. Several patterns were common to both data sets. However, the three-dimensional plot of the variables describing the structure of the arthropod community showed a notably rougher surface than in the case of Bornean beetles.
- 4 Although several factors may complicate the interpretation of the three-dimensional plots, long-term and continuous sampling may alter our perception of complex arthropod communities. This methodology is imperative for a proper understanding of arthropod community structure in rain forests.
170.
Patrick A. Garratt 《Environmental Biology of Fishes》1991,31(4):345-353
Synopsis The santerCheimerius nufar is widespread in tropical and subtropical waters of the western Indian Ocean and forms an important component of linefish catches along the east coast of southern Africa. Observations of spawning behaviour in captivity have revealed that spawning occurs during spring over a period of four months. Mating takes place at sunrise and may continue for up to 105 min (mean duration = 60 min). During spawning males become dark with prominent white markings. They become very aggressive and set up territories. Females remain a uniform silvery-pink. Mating occurs between males and females of similar sizes, culminating in egg and sperm release near the surface. Individual fish spawned up to 14 times during a morning. Streaking occurred throughout the season, with a second male joining a spawning pair and releasing gametes simultaneously. Slinger,Chrysoblephus puniceus, a dominant fish on offshore reefs in the same region, interfered with spawning throughout the season and were observed to eat eggs when they were released. The spawning strategy ofC. nufar is similar to protogynous species in several respects, indicating that this functional gonochorist may not conform to current theoretical predictions. 相似文献