全文获取类型
收费全文 | 517篇 |
免费 | 49篇 |
国内免费 | 1篇 |
出版年
2022年 | 8篇 |
2021年 | 12篇 |
2020年 | 5篇 |
2019年 | 4篇 |
2018年 | 11篇 |
2017年 | 10篇 |
2016年 | 9篇 |
2015年 | 16篇 |
2014年 | 30篇 |
2013年 | 29篇 |
2012年 | 41篇 |
2011年 | 16篇 |
2010年 | 28篇 |
2009年 | 29篇 |
2008年 | 34篇 |
2007年 | 21篇 |
2006年 | 28篇 |
2005年 | 24篇 |
2004年 | 27篇 |
2003年 | 19篇 |
2002年 | 10篇 |
2001年 | 13篇 |
2000年 | 11篇 |
1999年 | 13篇 |
1998年 | 6篇 |
1997年 | 5篇 |
1996年 | 8篇 |
1994年 | 2篇 |
1993年 | 3篇 |
1992年 | 7篇 |
1991年 | 7篇 |
1990年 | 14篇 |
1989年 | 6篇 |
1988年 | 9篇 |
1987年 | 6篇 |
1986年 | 5篇 |
1985年 | 6篇 |
1984年 | 4篇 |
1982年 | 6篇 |
1981年 | 2篇 |
1980年 | 2篇 |
1979年 | 3篇 |
1978年 | 4篇 |
1977年 | 1篇 |
1975年 | 1篇 |
1974年 | 2篇 |
1971年 | 4篇 |
1969年 | 1篇 |
1967年 | 2篇 |
1966年 | 1篇 |
排序方式: 共有567条查询结果,搜索用时 15 毫秒
131.
Z. Gattin S. Riniker P. J. Hore K. H. Mok W. F. van Gunsteren 《Protein science : a publication of the Protein Society》2009,18(10):2090-2099
The effects of temperature and urea denaturation (6M urea) on the dominant structures of the 20‐residue Trp‐cage mini‐protein TC5b are investigated by molecular dynamics simulations of the protein at different temperatures in aqueous and in 6M urea solution using explicit solvent degrees of freedom and the GROMOS force‐field parameter set 45A3. In aqueous solution at 278 K, TC5b is stable throughout the 20 ns of MD simulation and the trajectory structures largely agree with the NMR‐NOE atom–atom distance data available. Raising the temperature to 360 K and to 400 K, the protein denatures within 22 ns and 3 ns, showing that the denaturation temperature is well below 360 K using the GROMOS force field. This is 40–90 K lower than the denaturation temperatures observed in simulations using other much used protein force fields. As the experimental denaturation temperature is about 315 K, the GROMOS force field appears not to overstabilize TC5b, as other force fields and the use of continuum solvation models seem to do. This feature may directly stem from the GROMOS force‐field parameter calibration protocol, which primarily involves reproduction of condensed phase thermodynamic quantities such as energies, densities, and solvation free energies of small compounds representative for protein fragments. By adding 6M urea to the solution, the onset of denaturation is observed in the simulation, but is too slow to observe a particular side‐chain side‐chain contact (Trp6‐Ile4) that was experimentally observed to be characteristic for the denatured state. Interestingly, using temperature denaturation, the process is accelerated and the experimental data are reproduced. 相似文献
132.
Human interferon α-1 (hIFNA1) is one of several interferon α subtypes that have been studied and commercialized to treat various viral diseases including hepatitis B and C as well as malignant melanoma. Protein aggregation has been problematic for every step in commercial production, from purification to the packaging and delivery of pharmaceutical proteins. In a previous study, we demonstrated that a stabilizing peptide from the C-terminal acidic tail of α-synuclein (ATS) could be used as an effective fusion tag to increase the stability of target proteins such as human growth hormone (hGH) and granulocyte colony-stimulating factor (G-CSF). In this study, we applied this ATS fusion system to hIFNA1 in order to protect against the aggregation of hIFNA1 by environmental stresses, since hIFNA1 aggregates elicit an undesirable immune response in humans. As expected, ATS-fused hIFNA1 (hIFNA1-ATS) protein showed enhanced stability against thermal stress, agitation stress, and repetitive freeze/thawing stress in comparison with native hIFNA1. More importantly, hIFNA1-ATS fusion protein appeared to be 1.6 times more active than hIFNA1 in a cell anti-proliferation assay. Furthermore, the solubility of hIFNA1-ATS appeared to be 1.7 times higher than that of native protein. Our results suggest that the ATS-tag system could be a useful means for protecting hIFNA1 protein from aggregation by various external stresses and could be used to increase the solubility of protein. 相似文献
133.
Byoung Chan Kim Daniel Lopez‐Ferrer Sang‐Mok Lee Hye‐Kyung Ahn Sujith Nair Seong H. Kim Beom Soo Kim Konstantinos Petritis David G. Camp Jay W. Grate Richard D. Smith Yoon‐Mo Koo Man Bock Gu Professor Jungbae Kim Professor 《Proteomics》2009,9(7):1893-1900
A stable and robust trypsin‐based biocatalytic system was developed and demonstrated for proteomic applications. The system utilizes polymer nanofibers coated with trypsin aggregates for immobilized protease digestions. After covalently attaching an initial layer of trypsin to the polymer nanofibers, highly concentrated trypsin molecules are crosslinked to the layered trypsin by way of a glutaraldehyde treatment. This process produced a 300‐fold increase in trypsin activity compared with a conventional method for covalent trypsin immobilization, and proved to be robust in that it still maintained a high level of activity after a year of repeated recycling. This highly stable form of immobilized trypsin was resistant to autolysis, enabling repeated digestions of BSA over 40 days and successful peptide identification by LC‐MS/MS. This active and stable form of immobilized trypsin was successfully employed in the digestion of yeast proteome extract with high reproducibility and within shorter time than conventional protein digestion using solution phase trypsin. Finally, the immobilized trypsin was resistant to proteolysis when exposed to other enzymes (i.e., chymotrypsin), which makes it suitable for use in “real‐world” proteomic applications. Overall, the biocatalytic nanofibers with trypsin aggregate coatings proved to be an effective approach for repeated and automated protein digestion in proteomic analyses. 相似文献
134.
Chen L Li J Luo C Liu H Xu W Chen G Liew OW Zhu W Puah CM Shen X Jiang H 《Bioorganic & medicinal chemistry》2006,14(24):8295-8306
The 3C-like protease (3CLpro) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is one of the most promising targets for discovery of drugs against SARS, because of its critical role in the viral life cycle. In this study, a natural compound called quercetin-3-β-galactoside was identified as an inhibitor of the protease by molecular docking, SPR/FRET-based bioassays, and mutagenesis studies. Both molecular modeling and Q189A mutation revealed that Gln189 plays a key role in the binding. Furthermore, experimental evidence showed that the secondary structure and enzymatic activity of SARS-CoV 3CLpro were not affected by the Q189A mutation. With the help of molecular modeling, eight new derivatives of the natural product were designed and synthesized. Bioassay results reveal salient features of the structure–activity relationship of the new compounds: (1) removal of the 7-hydroxy group of the quercetin moiety decreases the bioactivity of the derivatives; (2) acetoxylation of the sugar moiety abolishes inhibitor action; (3) introduction of a large sugar substituent on 7-hydroxy of quercetin can be tolerated; (4) replacement of the galactose moiety with other sugars does not affect inhibitor potency. This study not only reveals a new class of compounds as potential drug leads against the SARS virus, but also provides a solid understanding of the mechanism of inhibition against the target enzyme. 相似文献
135.
Hugo YK Lam Philip M Kim Janine Mok Raffi Tonikian Sachdev S Sidhu Benjamin E Turk Michael Snyder Mark B Gerstein 《BMC bioinformatics》2010,11(1):243
Background
Many protein interactions, especially those involved in signaling, involve short linear motifs consisting of 5-10 amino acid residues that interact with modular protein domains such as the SH3 binding domains and the kinase catalytic domains. One straightforward way of identifying these interactions is by scanning for matches to the motif against all the sequences in a target proteome. However, predicting domain targets by motif sequence alone without considering other genomic and structural information has been shown to be lacking in accuracy. 相似文献136.
Park KJ Kang SJ Kim TM Lee YM Lee HC Song G Han JY 《In vitro cellular & developmental biology. Animal》2010,46(10):828-833
The production of chimeric birds is an important tool for the investigation of vertebrate development, the conservation of
endangered birds, and the development of various biotechnological applications. This study examined whether gamma (γ)-irradiation
depletes endogenous primordial germ cells and enhances the efficiency of somatic chimerism in chickens. An optimal irradiation
protocol for stage X embryos was determined after irradiation at various doses (0, 100, 300, 500, 600, 700, and 2,000 rad).
Exposure to 500 rad of γ-irradiation for 73 s significantly decreased the number of primordial germ cells (P < 0.0001). Somatic chimera hatchlings were then produced by transferring blastodermal cells from a Korean Oge into either
an irradiated (at 500 rad) or intact stage X White Leghorn embryo. An analysis of feather color pattern and polymerase chain
reaction-based species-specific amplification of various tissues of the hatchlings confirmed chimerism in most organs of the
chick produced from the irradiated recipient; a lesser degree of chimerism was observed in the non-irradiated control recipient.
In conclusion, the exposure of chick embryos to an optimized dose of γ-irradiation effectively depleted germ cells and yielded
greater somatic chimerism than non-irradiated control embryos. This technique can be applied to interspecies reproduction
or the production of transgenic birds. 相似文献
137.
138.
Human α-lactalbumin made lethal to tumor cells (HAMLET) and equine lysozyme with oleic acid (ELOA) are complexes consisting of protein and fatty acid that exhibit cytotoxic activities, drastically differing from the activity of their respective proteinaceous compounds. Since the discovery of HAMLET in the 1990s, a wealth of information has been accumulated, illuminating the structural, functional and therapeutic properties of protein complexes with oleic acid, which is summarized in this review. In vitro, both HAMLET and ELOA are produced by using ion-exchange columns preconditioned with oleic acid. However, the complex of human α-lactalbumin with oleic acid with the antitumor activity of HAMLET was found to be naturally present in the acidic fraction of human milk, where it was discovered by serendipity. Structural studies have shown that α-lactalbumin in HAMLET and lysozyme in ELOA are partially unfolded, 'molten-globule'-like, thereby rendering the complexes dynamic and in conformational exchange. HAMLET exists in the monomeric form, whereas ELOA mostly exists as oligomers and the fatty acid stoichiometry varies, with HAMLET holding an average of approximately five oleic acid molecules, whereas ELOA contains a considerably larger number (11- 48). Potent tumoricidal activity is found in both HAMLET and ELOA, and HAMLET has also shown strong potential as an antitumor drug in different in vivo animal models and clinical studies. The gain of new, beneficial function upon partial protein unfolding and fatty acid binding is a remarkable phenomenon, and may reflect a significant generic route of functional diversification of proteins via varying their conformational states and associated ligands. 相似文献
139.
Jobichen C Chakraborty S Li M Zheng J Joseph L Mok YK Leung KY Sivaraman J 《PloS one》2010,5(9):e12910
The recently identified type VI secretion system (T6SS) is implicated in the virulence of many Gram-negative bacteria. Edwardsiella tarda is an important cause of hemorrhagic septicemia in fish and also gastro- and extra-intestinal infections in humans. The E
. tarda
virulent protein (EVP) gene cluster encodes a conserved T6SS which contains 16 open reading frames. EvpC is one of the three major EVP secreted proteins and shares high sequence similarity with Hcp1, a key T6SS virulence factor from Pseudomonas aeruginosa. EvpC contributes to the virulence of E. tarda by playing an essential role in functional T6SS. Here, we report the crystal structure of EvpC from E. tarda PPD130/91 at a 2.8 Å resolution, along with functional studies of the protein. EvpC has a β-barrel domain with extended loops. The β-barrel consists of 11 anti-parallel β-strands with an α-helix located on one side. In solution, EvpC exists as a dimer at low concentration and as a hexamer at higher concentration. In the crystal, the symmetry related EvpC molecules form hexameric rings which stack together to form a tube similar to Hcp1. Structure based mutagenesis revealed that N-terminal negatively charged residues, Asp4, Glu15 and Glu26, and C-terminal positively charged residues, Lys161, Lys162 and Lys163, played crucial roles in the secretion of EvpC. Moreover, the localization study indicates the presence of wild type EvpC in cytoplasm, periplasm and secreted fractions, whereas the N-terminal and C-terminal mutants were found mostly in the periplasmic region and was completely absent in the secreted fraction. Results reported here provide insight into the structure, assembly and function of EvpC. Further, these findings can be extended to other EvpC homologs for understanding the mechanism of T6SS and targeting T6SS mediated virulence in Gram-negative pathogens. 相似文献
140.