全文获取类型
收费全文 | 1014篇 |
免费 | 68篇 |
国内免费 | 6篇 |
专业分类
1088篇 |
出版年
2022年 | 16篇 |
2021年 | 40篇 |
2020年 | 15篇 |
2019年 | 18篇 |
2018年 | 33篇 |
2017年 | 22篇 |
2016年 | 35篇 |
2015年 | 40篇 |
2014年 | 43篇 |
2013年 | 67篇 |
2012年 | 58篇 |
2011年 | 83篇 |
2010年 | 50篇 |
2009年 | 31篇 |
2008年 | 47篇 |
2007年 | 48篇 |
2006年 | 33篇 |
2005年 | 41篇 |
2004年 | 36篇 |
2003年 | 33篇 |
2002年 | 25篇 |
2001年 | 14篇 |
2000年 | 11篇 |
1999年 | 16篇 |
1998年 | 6篇 |
1997年 | 5篇 |
1996年 | 8篇 |
1995年 | 7篇 |
1994年 | 5篇 |
1993年 | 5篇 |
1992年 | 8篇 |
1991年 | 10篇 |
1990年 | 8篇 |
1989年 | 10篇 |
1987年 | 10篇 |
1986年 | 6篇 |
1985年 | 13篇 |
1984年 | 7篇 |
1983年 | 10篇 |
1982年 | 6篇 |
1981年 | 6篇 |
1980年 | 5篇 |
1979年 | 7篇 |
1978年 | 5篇 |
1977年 | 6篇 |
1976年 | 6篇 |
1975年 | 5篇 |
1974年 | 5篇 |
1971年 | 7篇 |
1969年 | 9篇 |
排序方式: 共有1088条查询结果,搜索用时 15 毫秒
41.
Probing the roles of two tryptophans surrounding the unique zinc coordination site in lipase family I.5 下载免费PDF全文
Emel Timucin Alexandra Cousido‐Siah André Mitschler Alberto Podjarny Osman Ugur Sezerman 《Proteins》2016,84(1):129-142
A unique zinc domain found in all of the identified members of the lipase family I.5 is surrounded by two conserved tryptophans (W61 and W212). In this study, we investigated the role of these hydrophobic residues in thermostability and thermoactivity of the lipase from Bacillus thermocatenulatus (BTL2) taken as the representative of the family. Circular dichroism spectroscopy revealed that the secondary structure of BTL2 is conserved by the tryptophan mutations (W61A, W212A, and W61A/W212A), and that W61 is located in a more rigid and less solvent exposed region than is W212. Thermal denaturation and optimal activity analyses pointed out that zinc induces thermostability and thermoactivity of BTL2, in which both tryptophans W61 and W212 play contributing roles. Molecular explanations describing the roles of these tryptophans were pursued by X‐ray crystallography of the open form of the W61A mutant and molecular dynamics simulations which highlighted a critical function for W212 in zinc binding to the coordination site. This study reflects the potential use of hydrophobic amino acids in vicinity of metal coordination sites in lipase biocatalysts design. Proteins 2016; 84:129–142. © 2015 Wiley Periodicals, Inc. 相似文献
42.
Calculation of the free energy of protein folding and delineation of its pre-organization are of foremost importance for understanding, predicting and designing biological macromolecules. Here, we introduce an energy smoothing variant of parallel tempering replica exchange Monte Carlo (REMS) that allows for efficient configurational sampling of flexible solutes under the conditions of molecular hydration. Its usage to calculate the thermal stability of a model globular protein, Trp cage TC5b, achieves excellent agreement with experimental measurements. We find that the stability of TC5b is attained through the coupled formation of local and non-local interactions. Remarkably, many of these structures persist at high temperature, concomitant with the origin of native-like configurations and mesostates in an otherwise macroscopically disordered unfolded state. Graph manifold learning reveals that the conversion of these mesostates to the native state is structurally heterogeneous, and that the cooperativity of their formation is encoded largely by the unfolded state ensemble. In all, these studies establish the extent of thermodynamic and structural pre-organization of folding of this model globular protein, and achieve the calculation of macromolecular stability ab initio, as required for ab initio structure prediction, genome annotation, and drug design. 相似文献
43.
44.
Osman H Mikes V Milat ML Ponchet M Marion D Prangé T Maume BF Vauthrin S Blein JP 《FEBS letters》2001,489(1):55-58
Cryptogein is a proteinaceous elicitor of plant defense reactions which also exhibits sterol carrier properties. In this study, we report that this protein binds fatty acids. The stoichiometry of the fatty acid-cryptogein complex is 1:1. Linoleic acid and dehydroergosterol compete for the same site, but elicitin affinity is 27 times lower for fatty acid than for sterol. We show that C7 to C12 saturated and C16 to C22 unsaturated fatty acids are the best ligands. The presence of double bonds markedly increases the affinity of cryptogein for fatty acids. A comparison between elicitins and known lipid transfer proteins is discussed. 相似文献
45.
Wei AC Ali MA Yoon YK Ismail R Choon TS Kumar RS Arumugam N Almansour AI Osman H 《Bioorganic & medicinal chemistry letters》2012,22(15):4930-4933
A series of twelve dispiropyrrolidines were synthesized using [3+2]-cycloaddition reactions. The synthesized compounds were screened for their antimycobacterial activity against M. tuberculosis H(37)Rv and INH resistant M. tuberculosis strains using agar dilution method, four of them showed good activity with MIC of less than 1 μM. Compound 4'-[5-(4-fluorophenyl)pyridin-3-yl]-1'-methyldispiro[indan-2,2' pyrrolidine-3',2″-indan]-1,3,1″-trione (4b) was found to be the most active with MIC of 0.1215 and 5.121 μM, respectively. 相似文献
46.
Cengiz?KayaEmail author Muhammed?Ashraf Osman?Sonmez Atilla?L.?Tuna Tahir?Polat Salih?Aydemir 《Acta Physiologiae Plantarum》2015,37(1):1741
The effects of thiamin (Thi) applied as seed soaking or foliar spray on some key physiological parameters were investigated in two differentially salt-responsive maize (Zea mays L.) cultivars, DK 5783 and Apex 836 F1, exposed to saline stress in two different experiments. An initial experiment (germination experiment) was designed to identify appropriate doses of Thi which could lessen the deleterious effects of salt on plants and screen all available maize cultivars for their differential tolerance to salt stress (100 mM NaCl). The seeds of nine maize cultivars were soaked for 24 h in solutions containing six levels of Thi (25, 50, 75, 100, 125 and 150 mg l?1). Based on the results obtained from the germination experiment, maize cultivar DK 5783 was found to be the most salt tolerant and Apex 836 as the most sensitive cultivar. Also, of six Thi levels used, two levels (100 and 125 mg l?1) were chosen for subsequent studies. In the second experiment (glasshouse experiment), two maize cultivars, DK 5783 (salt tolerant) and Apex 836 (salt sensitive) were subjected to saline regime (100 mM NaCl) and two levels of Thi (100 and 125 mg l?1) applied as foliar spray. Salt stress markedly suppressed shoot and root dry mass, total chlorophylls (“a” + “b”), leaf water potential and maximum fluorescence yield (Fv/Fm) in the plants of both maize cultivars, but it increased proline accumulation, leaf osmotic pressure, malondialdehyde (MDA) and hydrogen peroxide (H2O2) concentrations, electrolyte leakage (EL) as well as activities of some key antioxidant enzymes, superoxide dismutase (SOD; EC. 1.15.1.1), peroxidase (POD; EC. 1.11.1.7) and catalase (CAT; EC. 1.11.1.6). Salt-induced reduction in plant growth parameters was higher in the salt-sensitive cultivar, Apex 836, which was found to be associated with relatively increased EL, and MDA and H2O2 levels, and decreased activities of the key antioxidant enzymes. Application of Thi as seed soaking or foliar spray partly mitigated the deleterious effects of salinity on plants of both maize cultivars. The most promising effect of Thi on alleviation of adverse effects of salt stress on maize plants was found when it was applied as foliar spray at 100 mg l?1. Thiamin application considerably reduced tissue Na+ concentration, but improved those of N, P, Ca2+ and K+ in the salt-stressed maize plants. Exogenously applied thiamin-induced growth improvement in maize plants was found to be associated with reduced membrane permeability, MDA and H2O2 levels, and altered activities of some key antioxidant enzymes such as CAT, SOD and POD as well as increased photosynthetic pigment concentration under saline regime. 相似文献
47.
Johanne Mørch Jensen Fie C. SimonsenAmir Mastali Helle HaldIda Lillebro Frederik DinessLars Olsen Osman Mirza 《Peptides》2012
Proton-coupled oligopeptide transporters (POTs) utilize the electrochemical proton gradient to facilitate uptake of di- or tripeptide molecules. YjdL is one of four POTs found in Escherichia coli. It has shown an extraordinary preference for di- rather than tripeptides, and is therefore significantly different from prototypical POTs such as the human hPepT1. Nonetheless YjdL contains several highly conserved POT residues, which include Glu388 that is located in the putative substrate binding cavity. Here we present biophysical characterization of WT-YjdL and Glu388Gln. Isothermal titration calorimetrical studies exhibit a Kd of 14 μM for binding of Ala-Lys to WT-YjdL. Expectedly, no binding could be detected for the tripeptide Ala-Ala-Lys. Surprisingly however, binding could not be detected for Ala-Gln, although earlier studies indicated inhibitory potencies of Ala-Gln to be comparable to Ala-Lys (IC50 values of 0.6 compared to 0.3 mM). Finally, Ala-Lys binding to Glu388Gln was also undetectable which may support a previously suggested role in interaction with the ligand peptide N-terminus. 相似文献
48.
Molecular dynamics simulations of the 136 unique tetranucleotide sequences of DNA oligonucleotides. II: sequence context effects on the dynamical structures of the 10 unique dinucleotide steps 下载免费PDF全文
Dixit SB Beveridge DL Case DA Cheatham TE Giudice E Lankas F Lavery R Maddocks JH Osman R Sklenar H Thayer KM Varnai P 《Biophysical journal》2005,89(6):3721-3740
Molecular dynamics (MD) simulations including water and counterions on B-DNA oligomers containing all 136 unique tetranucleotide basepair steps are reported. The objective is to obtain the calculated dynamical structure for at least two copies of each case, use the results to examine issues with regard to convergence and dynamical stability of MD on DNA, and determine the significance of sequence context effects on all unique dinucleotide steps. This information is essential to understand sequence effects on DNA structure and has implications on diverse problems in the structural biology of DNA. Calculations were carried out on the 136 cases embedded in 39 DNA oligomers with repeating tetranucleotide sequences, capped on both ends by GC pairs and each having a total length of 15 nucleotide pairs. All simulations were carried out using a well-defined state-of-the-art MD protocol, the AMBER suite of programs, and the parm94 force field. In a previous article (Beveridge et al. 2004. Biophysical Journal. 87:3799-3813), the research design, details of the simulation protocol, and informatics issues were described. Preliminary results from 15 ns MD trajectories were presented for the d(CpG) step in all 10 unique sequence contexts. The results indicated the sequence context effects to be small for this step, but revealed that MD on DNA at this length of trajectory is subject to surprisingly persistent cooperative transitions of the sugar-phosphate backbone torsion angles alpha and gamma. In this article, we report detailed analysis of the entire trajectory database and occurrence of various conformational substates and its impact on studies of context effects. The analysis reveals a possible direct correspondence between the sequence-dependent dynamical tendencies of DNA structure and the tendency to undergo transitions that "trap" them in nonstandard conformational substates. The difference in mean of the observed basepair step helicoidal parameter distribution with different flanking sequence sometimes differs by as much as one standard deviation, indicating that the extent of sequence effects could be significant. The observations reveal that the impact of a flexible dinucleotide such as CpG could extend beyond the immediate basepair neighbors. The results in general provide new insight into MD on DNA and the sequence-dependent dynamical structural characteristics of DNA. 相似文献
49.
Makawi Ahmed Abdalhamid Osman Yu‐Jing Sun Rui‐Jia Li Hui Lin Dong‐Mei Zeng Xin‐Yu Chen Dongfang He Hui‐Wei Feng Zhao Yang Jin Wang Chaodong Wu Min Cui Jin‐Peng Sun Yuqing Huo Xiao Yu 《Journal of cellular and molecular medicine》2019,23(7):4653-4665
Severe reduction in the β‐cell number (collectively known as the β‐cell mass) contributes to the development of both type 1 and type 2 diabetes. Recent pharmacological studies have suggested that increased pancreatic β‐cell proliferation could be due to specific inhibition of adenosine kinase (ADK). However, genetic evidence for the function of pancreatic β‐cell ADK under physiological conditions or in a pathological context is still lacking. In this study, we crossed mice carrying LoxP‐flanked Adk gene with Ins2‐Cre mice to acquire pancreatic β ‐cell ADK deficiency (Ins2‐Cre±Adkfl/fl) mice. Our results revealed that Ins2‐Cre+/‐Adkfl/fl mice showed improved glucose metabolism and β‐cell mass in younger mice, but showed normal activity in adult mice. Moreover, Ins2‐Cre±Adkfl/fl mice were more resistant to streptozotocin (STZ) induced hyperglycaemia and pancreatic β‐cell damage in adult mice. In conclusion, we found that ADK negatively regulates β‐cell replication in young mice as well as under pathological conditions, such as STZ induced pancreatic β‐cell damage. Our study provided genetic evidence that specific inhibition of pancreatic β‐cell ADK has potential for anti‐diabetic therapy. 相似文献
50.