全文获取类型
收费全文 | 329篇 |
免费 | 12篇 |
国内免费 | 13篇 |
出版年
2024年 | 1篇 |
2023年 | 6篇 |
2021年 | 2篇 |
2020年 | 4篇 |
2019年 | 4篇 |
2018年 | 4篇 |
2017年 | 2篇 |
2016年 | 3篇 |
2015年 | 6篇 |
2014年 | 7篇 |
2013年 | 23篇 |
2012年 | 11篇 |
2011年 | 8篇 |
2010年 | 5篇 |
2009年 | 8篇 |
2008年 | 5篇 |
2007年 | 8篇 |
2006年 | 9篇 |
2005年 | 7篇 |
2004年 | 10篇 |
2003年 | 5篇 |
2002年 | 5篇 |
2001年 | 7篇 |
2000年 | 7篇 |
1999年 | 9篇 |
1998年 | 13篇 |
1997年 | 11篇 |
1996年 | 7篇 |
1995年 | 11篇 |
1994年 | 6篇 |
1993年 | 9篇 |
1992年 | 6篇 |
1991年 | 15篇 |
1990年 | 13篇 |
1989年 | 10篇 |
1988年 | 12篇 |
1987年 | 7篇 |
1986年 | 7篇 |
1985年 | 7篇 |
1984年 | 9篇 |
1983年 | 13篇 |
1982年 | 9篇 |
1981年 | 8篇 |
1980年 | 4篇 |
1979年 | 5篇 |
1978年 | 2篇 |
1977年 | 1篇 |
1976年 | 2篇 |
1973年 | 1篇 |
排序方式: 共有354条查询结果,搜索用时 218 毫秒
131.
Steve Boisvert David Joly Sébastien Leclerc Sridharan Govindachary Johanne Harnois Robert Carpentier 《Biometals》2007,20(6):879-889
The toxic effect of Ni2+ on photosynthetic electron transport was studied in a photosystem II submembrane fraction. It was shown that Ni2+ strongly inhibits oxygen evolution in the millimolar range of concentration. The inhibition was insensitive to NaCl but significantly
decreased in the presence of CaCl2. Maximal chlorophyll fluorescence, together with variable fluorescence, maximal quantum yield of photosystem II, and flash-induced
fluorescence decays were all significantly declined by Ni2+. Further, the extrinsic polypeptides of 16 and 24 kDa associated with the oxygen-evolving complex of photosystem II were
depleted following Ni2+ treatment. It was deduced that interaction of Ni2+ with these polypeptides caused a conformational change that induced their release together with Ca2+ from the oxygen-evolving complex of photosystem II with consequent inhibition of the electron transport activity. 相似文献
132.
Günter Schwarzmann Bernadette Breiden Konrad Sandhoff 《Journal of lipid research》2015,56(10):1861-1879
A Förster resonance energy transfer-based fusion and transfer assay was developed to study, in model membranes, protein-mediated membrane fusion and intermembrane lipid transfer of fluorescent sphingolipid analogs. For this assay, it became necessary to apply labeled reporter molecules that are resistant to spontaneous as well as protein-mediated intermembrane transfer. The novelty of this assay is the use of nonextractable fluorescent membrane-spanning bipolar lipids. Starting from the tetraether lipid caldarchaeol, we synthesized fluorescent analogs with fluorophores at both polar ends. In addition, we synthesized radioactive glycosylated caldarchaeols. These labeled lipids were shown to stretch through bilayer membranes rather than to loop within a single lipid layer of liposomes. More important, the membrane-spanning lipids (MSLs) in contrast to phosphoglycerides proved to be nonextractable by proteins. We could show that the GM2 activator protein (GM2AP) is promiscuous with respect to glycero- and sphingolipid transfer. Saposin (Sap) B also transferred sphingolipids albeit with kinetics different from GM2AP. In addition, we could unambiguously show that the recombinant activator protein Sap C x His6 induced membrane fusion rather than intermembrane lipid transfer. These findings showed that these novel MSLs, in contrast with fluorescent phosphoglycerolipids, are well suited for an uncompromised monitoring of membrane fusion and intermembrane lipid transfer. 相似文献
133.
Sarah R. MacEwan Wafa Hassouneh Ashutosh Chilkoti 《Journal of visualized experiments : JoVE》2014,(88)
Elastin-like polypeptides are repetitive biopolymers that exhibit a lower critical solution temperature phase transition behavior, existing as soluble unimers below a characteristic transition temperature and aggregating into micron-scale coacervates above their transition temperature. The design of elastin-like polypeptides at the genetic level permits precise control of their sequence and length, which dictates their thermal properties. Elastin-like polypeptides are used in a variety of applications including biosensing, tissue engineering, and drug delivery, where the transition temperature and biopolymer architecture of the ELP can be tuned for the specific application of interest. Furthermore, the lower critical solution temperature phase transition behavior of elastin-like polypeptides allows their purification by their thermal response, such that their selective coacervation and resolubilization allows the removal of both soluble and insoluble contaminants following expression in Escherichia coli. This approach can be used for the purification of elastin-like polypeptides alone or as a purification tool for peptide or protein fusions where recombinant peptides or proteins genetically appended to elastin-like polypeptide tags can be purified without chromatography. This protocol describes the purification of elastin-like polypeptides and their peptide or protein fusions and discusses basic characterization techniques to assess the thermal behavior of pure elastin-like polypeptide products. 相似文献
134.
Maize and Arabidopsis thaliana class 1 reversibly glycosylated polypeptides (C1RGPs) are plasmodesmata-associated proteins. Previously, overexpression of Arabidopsis C1RGP AtRGP2 in Nicotiana tabacum was shown to reduce intercellular transport of photoassimilate, resulting in stunted, chlorotic plants, and inhibition of local cell-to-cell spread of tobacco mosaic virus (TMV). Here, we used virus induced gene silencing to examine the effects of reduced levels of C1RGPs in Nicotiana benthamiana. Silenced plants show wild-type growth and development. Intercellular transport in silenced plants was probed using fluorescently labeled TMV and its movement protein, P30. P30 shows increased cell-to-cell movement and TMV exhibited accelerated systemic spread compared with control plants. These results support the hypothesis that C1RGPs act to regulate intercellular transport via plasmodesmata. 相似文献
135.
Sortagging is a versatile method for site‐specific modification of proteins as applied to a variety of in vitro reactions. Here, we explore possibilities of adapting the sortase method for use in living cells. For intracellular sortagging, we employ the Ca2+‐independent sortase A transpeptidase (SrtA) from Streptococcus pyogenes. Substrate proteins were equipped with the C‐terminal sortase‐recognition motif (LPXTG); we used proteins with an N‐terminal (oligo)glycine as nucleophiles. We show that sortase‐dependent protein ligation can be achieved in Saccharomyces cerevisiae and in mammalian HEK293T cells, both in the cytosol and in the lumen of the endoplasmic reticulum (ER). ER luminal sortagging enables secretion of the reaction products, among which circular polypeptides. Protein ligation of substrate and nucleophile occurs within 30 min of translation. The versatility of the method is shown by protein ligation of multiple substrates with green fluorescent protein‐based nucleophiles in different intracellular compartments. 相似文献
136.
Song Y McFarland DC Velleman SG 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2012,161(2):271-278
Syndecan-4 is composed of a core protein and covalently attached glycosaminoglycan (GAG) and N-linked glycosylated (N-glycosylated) chains. The core protein is divided into extracellular, transmembrane, and cytoplasmic domains. The cytoplasmic domain has two conserved regions and a variable region in the middle. The Ser residue in the conserved region 1 and the Tyr residue in the variable region are important in regulating protein kinase C alpha (PKCα) membrane localization and focal adhesion formation. The objective of the current study was to investigate the role of syndecan-4 Ser and Tyr residues in combination with the GAG and N-glycosylated chains in turkey satellite cell proliferation, differentiation, fibroblast growth factor 2 (FGF2) responsiveness, and PKCα membrane localization. Site-directed mutagenesis was used to generate Ser and Tyr mutants with or without GAG and N-glycosylated chains. The wild type and mutant syndecan-4 constructs were transfected into turkey satellite cells. The over-expression of Ser and Tyr mutants increased cell proliferation and differentiation and decreased membrane localization of PKCα. Furthermore, Ser mutants enhanced cellular responsiveness to FGF2. The results from this study are the first demonstration of a role of syndecan-4 cytoplasmic domain Ser and Tyr residues in regulating satellite cell proliferation, differentiation, and the modulation of cellular responsiveness to FGF2. 相似文献
137.
A new isolation procedure for Kalata polypeptides from the tropical plant Oldenlandia affinis DC is described. Fractions were screened by thin-layer chromatography, and Van Urk positive fractions were tested for oxytocic activity in estrogenized rat uteri. By using this procedure, we were able to isolate and characterize three macrocyclic polypeptides with uterine activity. Their amino acid sequence and biological effects have been analyzed, and their NMR spectra were compared with those of the earlier ones. All three peptides showed hemolytic activity on human blood, and were tested for antibiotic effect against E. coli, Staphylococcus aureus, and Hemophilus influenzae. 相似文献
138.
Kenji Shibata Toshiyuki Suzawa Tetsuji Ohno Koji Yamada Takeo Tanaka Eiji Tsukuda Yuzuru Matsuda Motoo Yamasaki 《Bioorganic & medicinal chemistry》1998,6(12):2459-2467
Hybrid peptides were constructed from endothelin B receptor (ETB) selective antagonist RES-701-1 (1) and endothelin (ET-1). They have N-terminal 10 amino acids derived from 1 and C-terminal 10 amino acids derived from ET-1. RES-701-1(1-10)-[Ala15]ET-1(12-21) and its analogues substituted or truncated at the residues derived from RES-701-1 had proved to possess high receptor binding activity selective for ETB as well as 1. Substitutions at the residues derived from ET-1 had produced some analogues that possessed high affinity not only for ETB but for ETA. Although all analogues had antagonistic effects on ETA, some analogues had proved to function as agonist on ETB confirmed by the changes in intracellular calcium concentrations of ET receptor-transfected COS-7 cells. We have found four types of ET receptor-binding peptides: (1) ETB-selective agonist with weak ETA antagonism (3, KT7421); (2) ETB-selective antagonist with weak ETA antagonism (29, KT7539); (3) ETB agonist with potent ETA antagonism (27, KT7538); and (4) non-selective ETA/ETB antagonist (26, KT7540). 相似文献
139.
动物类中药的有效成分以蛋白多肽为主,因此活性蛋白多肽具有重要的医疗保健价值。文章分析了沉淀法、色谱法、膜分离法以及电泳法的基本原理和主要适用范围,综述了这些方法在动物源活性蛋白多肽的分离纯化中的应用,为动物源蛋白多肽的分离纯化与进一步研究提供参考,以期开发出高效、经济和环保的蛋白多肽分离纯化新技术。 相似文献
140.
Shah M Hsueh PY Sun G Chang HY Janib SM MacKay JA 《Protein science : a publication of the Protein Society》2012,21(6):743-750
Protein polymers are repetitive polypeptides produced by ribosomal biosynthetic pathways; furthermore, they offer emerging opportunities in drug and biopharmaceutical delivery. As for any polymer, biodegradation is one of the most important determinants affecting how a protein polymer can be utilized in the body. This study was designed to characterize the proteolytic biodegradation for a library of protein polymers derived from the human tropoelastin, the Elastin-like polypeptides (ELPs). ELPs are of particular interest for controlled drug delivery because they reversibly transition from soluble to insoluble above an inverse phase transition temperature (T(t)). More recently, ELP block copolymers have been developed that can assemble into micelles; however, it remains unclear if proteases can act on these ELP nanoparticles. For the first time, we demonstrate that ELP nanoparticles can be degraded by two model proteases and that comparable proteolysis occurs after cell uptake into a transformed culture of murine hepatocytes. Both elastase and collagenase endopeptidases can proteolytically degrade soluble ELPs. To our surprise, the ELP phase transition was protective against collagenase but not to elastase activity. These findings enhance our ability to predict how ELPs will biodegrade in different physiological microenvironments and are essential to develop protein polymers into biopharmaceuticals. 相似文献