全文获取类型
收费全文 | 330篇 |
免费 | 5篇 |
国内免费 | 18篇 |
出版年
2021年 | 2篇 |
2019年 | 3篇 |
2018年 | 3篇 |
2017年 | 3篇 |
2016年 | 3篇 |
2015年 | 10篇 |
2014年 | 13篇 |
2013年 | 18篇 |
2012年 | 14篇 |
2011年 | 23篇 |
2010年 | 9篇 |
2009年 | 14篇 |
2008年 | 25篇 |
2007年 | 17篇 |
2006年 | 19篇 |
2005年 | 11篇 |
2004年 | 18篇 |
2003年 | 10篇 |
2002年 | 8篇 |
2001年 | 2篇 |
2000年 | 4篇 |
1999年 | 6篇 |
1998年 | 7篇 |
1997年 | 5篇 |
1996年 | 7篇 |
1995年 | 8篇 |
1994年 | 6篇 |
1993年 | 1篇 |
1992年 | 7篇 |
1991年 | 7篇 |
1990年 | 1篇 |
1989年 | 5篇 |
1988年 | 5篇 |
1987年 | 3篇 |
1986年 | 4篇 |
1985年 | 4篇 |
1984年 | 12篇 |
1983年 | 3篇 |
1982年 | 8篇 |
1981年 | 5篇 |
1980年 | 7篇 |
1979年 | 2篇 |
1978年 | 1篇 |
1976年 | 5篇 |
1974年 | 2篇 |
1972年 | 2篇 |
1971年 | 1篇 |
排序方式: 共有353条查询结果,搜索用时 15 毫秒
71.
Zabłotna E Dysasz H Lesner A Jaśkiewicz A Kaźmierczak K Miecznikowska H Rolka K 《Biochemical and biophysical research communications》2004,319(1):185-188
A tetrapeptide combinatorial library, considered as chromogenic substrates of bovine beta-trypsin, was synthesized by the solid phase method. The peptides contain an analog of p-nitroanilide, obtained by attaching 5-amino-2-nitrobenzoic acid (Anb(5,2)) to the C-termini. Deconvolution of the peptide library, performed in solution using an iterative method, yielded four efficient trypsin substrates. The most active one, Phe-Val-Pro-Arg-Anb(5,2)-NH(2), appeared to be 125-fold more active than Bz-D,L-Arg-pNA (BAPNA) used as a reference compound. The reported method of designing trypsin chromogenic substrate libraries is straightforward. Such p-nitroanilides may be useful for the investigation of any protease substrate specificity. 相似文献
72.
Cyanopeptolin S, a sulfate-containing depsipeptide from a water bloom of Microcystis sp. 总被引:1,自引:0,他引:1
Clemens Jakobi Lukas Oberer Charles Quiquerez Wilfried A. König Jürgen Weckesser 《FEMS microbiology letters》1995,129(2-3):129-133
Abstract A new sulfated, cyclic depsipeptide, called cyanopeptolin S, from Microcystis sp. was isolated from a water bloom in the Auensee/Leipzig (Germany). The depsipeptide had a relative molecular mass of 925 and contained l-arginine, l-threonine, l-isoleucine, N-methyl-l-phenylalanine, a l-glutamic acid-δ-aldehyde ring system and a sulfated d-configurated glyceric acid as a side chain. The structure was elucidated by means of two-dimensional 1 H and 13 C nuclear magnetic resonance spectroscopy, fast atom bombardment mass spectroscopy, Fourier transformed infrared spectroscopy and combined gas-liquid chromatography/mass spectrometry. Cyanopeptolin S inhibited trypsin with an IC50 ≤ 0.2 μg ml−1 . 相似文献
73.
Kinetic study of thermal inactivation for native and methoxypolyethylene glycol modified trypsin 总被引:1,自引:0,他引:1
Bovine pancreatic trypsin (Ti) has been modified with four kinds of methoxypolyethylene glycol (MPEG, molecular masses 350, 750, 2000 and 5000 Da) to enhance thermostability. The MPEG-modified Ti was more stable against temperature than the native form, the larger molecular mass moiety of MPEG showing higher thermostabillty. To investigate the mechanism of thermal inactivation, a new kinetic model, which has the ability of taking the thermal denaturation and autolysis effects of the proteases into account, has been used to analyze the thermal inactivation process of the native and modified Ti in detail. The kinetic analysis showed that the stabilization effect caused by MPEG modification was the result of a decrease in autolysis rate and a decrease in the rate of thermal denaturation. In addition, the possible mechanism of reduced autolysis and lower thermal denaturation rate were also discussed. 相似文献
74.
Umesh C. Haldar Sanat K. Saha Ronald C. Beavis Nirmal K. Sinha 《Journal of Protein Chemistry》1996,15(2):177-184
Two trypsin inhibitors, LA-1 and LA-2, have been isolated from ridged gourd (Luffa acutangula Linn.) seeds and purified to homogeneity by gel filtration followed by ion-exchange chromatography. The isoelectric point is atpH 4.55 for LA-1 and atpH 5.85 for LA-2. The Stokes radius of each inhibitor is 11.4 å. The fluorescence emission spectrum of each inhibitor is similar to that of the free tyrosine. The biomolecular rate constant of acrylamide quenching is 1.0×109 M–1 sec–1 for LA-1 and 0.8 × 109 M–1 sec–1 for LA-2 and that of K2HPO4 quenching is 1.6×1011 M–1 sec–1 for LA-1 and 1.2×1011M–1 sec–1 for LA-2. Analysis of the circular dichroic spectra yields 40%-helix and 60%-turn for La-1 and 45%-helix and 55%-turn for LA-2. Inhibitors LA-1 and LA-2 consist of 28 and 29 amino acid residues, respectively. They lack threonine, alanine, valine, and tryptophan. Both inhibitors strongly inhibit trypsin by forming enzymeinhibitor complexes at a molar ratio of unity. A chemical modification study suggests the involvement of arginine of LA-1 and lysine of LA-2 in their reactive sites. The inhibitors are very similar in their amino acid sequences, and show sequence homology with other squash family inhibitors. 相似文献
75.
Elementary Na+ currents were recorded at 19°C in inside-out patches from cultured neonatal rat cardiocytes. In analyzing the sensitivity of chemically modified Na+ channels to several class 1 antiarrhythmic drugs, the hypothesis was tested that removal of Na+ inactivation may be accompanied by a distinct responsiveness to these drugs, open channel blockade.Iodate-modified and trypsin-modified cardiac Na+ channels are noninactivating but strikingly differ from each other by their open state kinetics, a O1–O2 reaction (open(1) 1.4±0.3 msec; open(2) 5.4±1.1 msec; at –40 mV) in the former and a single open state (open 3.0±0.5 msec; at –40 mV) in the latter. Lidocaine (150 mol/liter) like propafenone (10 mol/liter), diprafenone (10 mol/liter) and quinidine (20 mol/liter) in cytoplasmic concentrations effective to depress NP
o
significantly can interact with both types of noninactivating Na+ channels to reduce the dwell time in the conducting configuration. lodate-modified Na+ channels became drug sensitive during the O2 state. At –40 mV, for example, lidocaine reduced open(2) to 62±5% of the control without detectable changes in open(1). No evidence could be obtained that these inhibitory molecules would flicker-block the open Na+ pore. Drug-induced shortening of the open state, thus, is indicative for a distinct mode of drug action, namely interference with the gating process. Lidocaine proved less effective to reduce open(2) when compared with the action of diprafenone. Both drugs apparently interacted with individual association rate constants, alidocaine was 0.64×106 mol–1 sec–1 and adiprafenone 13.6×106 mol–1 sec–1. Trypsin-modified Na+ channels also appear capable of discriminating among these antiarrhythmics, the ratio adiprafenone/alidocaine even exceeded the value in iodate-modified Na+ channels. Obviously, this antiarrhythmic drug interaction with chemically modified Na+ channels is receptor mediated: drug occupation of such a hypothetical hidden receptor that is not available in normal Na+ channels may facilitate the exit from the open state.This work was supported by a grant of the Deutsche Forschungsgemeinschaft (Ko 778/2-4), Bonn. 相似文献
76.
K. P. Kollipara L. Singh T. Hymowitz 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(8):986-993
Variation in the trypsin inhibitors (TIs) and the chymotrypsin inhibitors (CIs) among 69 pigeonpea [Cajanus cajan (L.) Millsp.] strains from a wide geographical distribution and among 17 accessions representing seven wild Cajanus species was studied by electrophoretic banding pattern comparisons and by spectrophotometric activity assays. The TI and CI electrophoretic migration patterns among the pigeonpea strains were highly uniform but varied in the inhibitor band intensities. The migration patterns of the inhibitors in the wild Cajanus species were highly species specific. The mean TI activity of pigeonpea strains (2279 units) was significantly higher than that of the wild Cajanus species (1407 units). However, the mean CI activity in the pigeonpea strains (62 units) was much lower than that in the wild species (162 units). Kenya 2 and ICP 9151 were the lowest and the highest, respectively, in both the TI and CI activities among all the pigeonpea strains used in this study. A highly-significant positive correlation was observed between the TI and CI activities. The Bowman-Birk type inhibitors with both TI and CI activities were identified in all the pigeonpea strains and also in the accessions of all the wild species except C. volubilis (Blanco) Blanco. The C. volubilis accession ICPW 169 was found to be null for both CI bands and CI activity. Environment, strain, and environment x strain interaction showed highly-significant effects on both the TI and CI activities. Growing the pigeonpea strains at a different environment from their area of adaptation increased TI and CI activities and also altered the maturity period. 相似文献
77.
Purification and characterization of a trypsin inhibitor from Putranjiva roxburghii seeds 总被引:1,自引:0,他引:1
Chaudhary NS Shee C Islam A Ahmad F Yernool D Kumar P Sharma AK 《Phytochemistry》2008,69(11):2120-2126
A highly stable and potent trypsin inhibitor was purified to homogeneity from the seeds of Putranjiva roxburghii belonging to Euphorbiaceae family by acid precipitation, cation-exchange and anion-exchange chromatography. SDS-PAGE analysis, under reducing condition, showed that protein consists of a single polypeptide chain with molecular mass of approximately 34 kDa. The purified inhibitor inhibited bovine trypsin in 1:1 molar ratio. Kinetic studies showed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 1.4x10(-11) M. The inhibitor retained the inhibitory activity over a broad range of pH (pH 2-12), temperature (20-80 degrees C) and in DTT (up to100 mM). The complete loss of inhibitory activity was observed above 90 degrees C. CD studies, at increasing temperatures, demonstrated the structural stability of inhibitor at high temperatures. The polypeptide backbone folding was retained up to 80 degrees C. The CD spectra of inhibitor at room temperature exhibited an alpha, beta pattern. N-terminal amino acid sequence of 10 residues did not show any similarities to known serine proteinase inhibitors, however, two peptides obtained by internal partial sequencing showed significant resemblance to Kunitz-type inhibitors. 相似文献
78.
Proteases are implicated in various diseases and several have been identified as potential drug targets or biomarkers. As a result, protease activity assays that can be performed in high throughput are essential for the screening of inhibitors in drug discovery programs. Here we describe the development of a simple, general method for the characterization of protease activity and its use for inhibitor screening. GFP was genetically fused to a comparatively unstable Tus protein through an interdomain linker containing a specially designed protease site, which can be proteolyzed. When this Tus–GFP fusion protein substrate is proteolyzed it releases GFP, which remains in solution after a short heat denaturation and centrifugation step used to eliminate uncleaved Tus–GFP. Thus, the increase in GFP fluorescence is directly proportional to protease activity. We validated the protease activity assay with three different proteases, i.e., trypsin, caspase 3, and neutrophil elastase, and demonstrated that it can be used to determine protease activity and the effect of inhibitors with small sample volumes in just a few simple steps using a fluorescence plate reader. 相似文献
79.
In-gel digestion has been standardised using a poly(propylene) disposable. We designed a four-step rapid and simple in-gel digestion protocol which is carried out in one self-contained reaction tube avoiding keratin contamination. In order to quantify the efficiency of in-gel digestion, we developed a rapid on-column peptide acetylation protocol. Results show that trypsin in-gel uptake is increased and in-gel digestion is 90% complete within 15 min. We further show that spectrum quality, peptide yield and sequence coverage for mass spectrometric analysis are enhanced. We utilise 2-D PAGE separation of photosystem II from barley to demonstrate that the protocol facilitates identification of highly hydrophobic membrane proteins. 相似文献
80.
Sales MP Andrade LB Ary MB Miranda MR Teixeira FM Oliveira AS Fernandes KV Xavier-Filho J 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2005,142(4):422-426
Callosobruchus maculatus (Cm) and Zabrotes subfasciatus (Zs) were reared on resistant (IT81D-1045) and on susceptible (Epace 10) cowpea seeds. The emergence of adult insects, total developmental period (TDP) and excretion of trypsin inhibitor and vicilin were determined for both bruchid populations. Parameter evaluation showed that the Zs populations emerged from both seeds had no significant differences in emergence and TDP. The Cm population raised from resistant seeds had lower emergence (5.6+/-1.3%) and delayed TDP (46+/-1.25 days) than those emerged from susceptible seeds. The excretion of defense proteins showed that Zs reared in resistant seeds excreted 1.7 times more trypsin inhibitor, but this did not affect emergence or TDP. Furthermore, Cm population emerged from resistant seeds excreted 7 times higher vicilin and 0.4 times less trypsin inhibitor than that emerged from susceptible seeds. These results indicate that vicilins from resistant seeds are involved to significantly longer TDP (46 days) and also drastic reduction of insect emergence ( approximately 5%) of C. maculatus. 相似文献