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1.
A.Q.H. Habets-Crützen S.J.N. Carlier J.A.M. de Bont D. Wistuba V. Schurig S. Hartmans J. Tramper 《Enzyme and microbial technology》1985,7(1):17-21
Resting cells of ethene grown Mycobacterium 2W produced 1,2-epoxypropane stereospecifically from propene as revealed by optical rotation, 1H n.m.r. using a chiral shift reagent, and also by complexation gas chromatography involving a glass capillary column coated with an optically active metal chelate. The gas-liquid chromatography method allowed the rapid screening of 11 strains with regard to stereospecific formation of 1,2-epoxypropane, 1,2-epoxybutane and 1-chloro-2,3-epoxypropane. Bacteria grown on either ethene, propene or butadiene all predominantly produced the R form of 1,2-epoxypropane from propene and 1,2-epoxybutane from 1-butene while the strains tested for 1-chloro-2,3-epoxypropane production from 3-chloro-1-propene predominantly accumulated the S enantiomer. 相似文献
2.
Competition between Ca2+ and Mg2+ for binding to a single high affinity site on actin has been confirmed. Occupancy of this site only by either Ca2+ or Mg2+ affects the conformation of actin and its ability to form nuclei and hydrolyze ATP. G-actin binds the beta gamma-bidentate CrATP, a substitution inert analog of metal-ATP complexes, and shows a high specificity for the lambda isomers. Binding of CrATP to ADP-actin is accompanied by the dissociation of tightly bound ADP and Ca2+. CrATP-actin shows a high tendency to form nuclei, like MgATP-actin. Polymerization of CrATP-actin is accompanied by cleavage of the gamma-phosphate, but subsequent Pi release cannot occur because the product of the reaction is the stable CrADP-Pi complex. All these results support the view that the divalent metal ion tightly bound to actin interacts with the beta- and gamma-phosphates of ATP in the nucleotide site. 相似文献
3.
M Van Troys D Dewitte M Goethals M F Carlier J Vandekerckhove C Ampe 《The EMBO journal》1996,15(2):201-210
We characterized in detail the actin binding site of the small actin-sequestering protein thymosin beta 4 (T beta 4) using chemically synthesized full-length T beta 4 variants. The N-terminal part (residues 1-16) and a hexapeptide motif (residues 17-22) form separate structural entities. In both, we identified charged and hydrophobic residues that participate in the actin interaction using chemical cross-linking, complex formation in native gels and actin-sequestering experiments. Quantitative data on the activity of the variants and circular dichroism experiments allow to present a model in which the N-terminal part needs to adopt an alpha-helix for actin binding and interacts through a patch of hydrophobic residues (6M-I-F12) on one side of this helix. Also, electrostatic contacts between actin and lysine residues 18, in the motif, and 14, in the N-terminal alpha-helix, appear important for binding. The residues critical for contacting actin are conserved throughout the beta-thymosin family and in addition to this we identify a similar pattern in the C-terminal headpiece of villin and dematin. 相似文献
4.
D el Allaf S Cremers V D'Orio J Carlier 《Archives internationales de physiologie et de biochimie》1984,92(4):S49-S55
The haemodynamic effects of an optimal dose of dobutamine (DUo) (6.7 +/- 4.2 micrograms kg-1 min-1) and the combination of this optimal dose minus 2.5 micrograms kg-1 min-1 of dobutamine (DU) plus dopamine 2.5 micrograms kg-1 min-1 (DA) were studied in a first group of 12 consecutive patients with acute myocardial infarction (AMI) and cardiac failure (CF). DUo decreased pulmonary wedge pressure from 23.5 to 16 mm Hg (P less than 0.01), systemic vascular resistance from 1 774 to 1 417 dynes s cm-5 (P less than 0.01). DUo increased cardiac output from 3.21 to 4.55 litres/min (P less than 0.01) and urinary flow (UF) from 20 to 68 ml/h (P less than 0.01). Heart rate and blood pressure did not change significantly. DUo - DU + DA significantly increased UF from 68 to 107 ml/h (P less than 0.05) while the other parameters remained unchanged with respect to DUo. The positive effect of DA on UF was confirmed in a second group of 12 consecutive patients by comparing the successive effects of DA + DUo and DUo + DU : all previously described parameters remained unchanged except UF which decreased from 107 to 65 (P less than 0.01). We conclude that in patients with CF and AMI, association of DA and DUo is useful in obtaining both inotropic and diuretic effects. 相似文献
5.
Chromium GTP (CrGTP) has been used to probe the stereochemistry of metal-GTP binding to exchangeable site of tubulin and to examine the fate and role of nucleotide-bound metal ion in GTP hydrolysis associated with microtubule assembly. The absolute stereoconfiguration of the two pairs of diastereomers of beta,gamma-bidentate CrGTP has been determined by comparison of their visible circular dichroism spectra with those of the beta,gamma-CrATP isomers whose configurations have been established (Lin, I., and Dunaway-Mariano, D. (1988) J. Am. Chem. Soc. 110, 950-956). Tubulin binds metal-GTP preferentially in the delta pseudoaxial configuration. CrGTP-tubulin shows a high propensity to undergo tubulin-tubulin interactions with associated hydrolysis of CrGTP. Hydrolysis of CrGTP in microtubule assembly develops in two consecutive steps: cleavage of the gamma-phosphate followed by release of Pi and chromium. In contrast to other NTPases (actin, hexokinase) tubulin appears able to catalyze the dissociation of the stable chromium-phosphate bonds, which implies a highly nucleophilic environment of the binding site of the metal-triphosphate moiety of GTP. Microtubules assembled from CrGTP-tubulin are made of 90% GDP subunits, and their stability is linked to a 10% proportion of CrGDP-Pi subunits, scattered along the microtubule, from which Pi does not dissociate. The possibility is evoked that some tubulin variants do not catalyze release of Pi and metal ion efficiently, and their presence could affect microtubule dynamics. 相似文献
6.
J. M. Thevelein J. A. van Assche K. Heremans S. Y. Gerlsma A. R. Carlier 《Antonie van Leeuwenhoek》1981,47(5):393-404
The heat activation of trehalase in extracts of sporangiospores of Phycomyces blakesleeanus, following the induction of germination by heat activation and the gelatinization of potato starch granules were studied under different conditions in order to discriminate between several phenomena as possible triggers in the activation of trehalase.Short-chain alcohols (from methanol to pentanol) lower the activation temperature of trehalase while long-chain alcohols (from heptanol to nonanol) raise it. Short-chain alcohols also lower the gelatinization temperature of potato starch granules, while long-chain alcohols, hexanol and heptanol have hardly any influence on the gelatinization temperature. Octanol raises the gelatinization temperature. More polar phenols lower the activation temperature of trehalase, while more apolar phenols will raise it. The gelatinization temperature of starch granules is more lowered by the polar polyphenols than by the more apolar phenols.The effect of high pressure on starch gelatinization was investigated in order to compare data from such a model system with the data on trehalase activation.The gelatinization temperature of starch granules is shifted upwards with about 3–5 K/1000 atm (1.013×105 kPa). Pressures higher than 1500 atm do not further increase the gelatinization temperature. However, no reversal of the effect, as occurs with protein conformational changes, is seen with pressure up to 2500 atm. Also for trehalase activation we find a continuous upward shift of the activation temperature with about 5–9 K/1000 atm. These data are in agreement with a thermal transition in a polysaccharide matrix, being the trigger in the heat activation of trehalase. 相似文献
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9.
Carlier L Couprie J le Maire A Guilhaudis L Milazzo-Segalas I Courçon M Moutiez M Gondry M Davoust D Gilquin B Zinn-Justin S 《Protein science : a publication of the Protein Society》2007,16(12):2750-2755
Human KIN17 is a 45-kDa eukaryotic DNA- and RNA-binding protein that plays an important role in nuclear metabolism and in particular in the general response to genotoxics. Its amino acids sequence contains a zinc finger motif (residues 28-50) within a 30-kDa N-terminal region conserved from yeast to human, and a 15-kDa C-terminal tandem of SH3-like subdomains (residues 268-393) only found in higher eukaryotes. Here we report the solution structure of the region 51-160 of human KIN17. We show that this fragment folds into a three-alpha-helix bundle packed against a three-stranded beta-sheet. It belongs to the winged helix (WH) family. Structural comparison with analogous WH domains reveals that KIN17 WH module presents an additional and highly conserved 3(10)-helix. Moreover, KIN17 WH helix H3 is not positively charged as in classical DNA-binding WH domains. Thus, human KIN17 region 51-160 might rather be involved in protein-protein interaction through its conserved surface centered on the 3(10)-helix. 相似文献
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