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111.
J. R. Albani 《Journal of fluorescence》1998,8(3):213-224
The present work reports for the first time the development of a method that allowed us to obtain crystals of orosomucoid complexed to progesterone. Then we investigated the dynamics of the microenvironments of the two buried Trp residues in the crystals of protein, by the red-edge excitation spectra method. The fluorescence excitation spectrum of the crystals is characteristic of that known for Trp residues (max = 290 nm and bandwidth = 38 ± 1 nm), indicating that the Trp residues are responsible for the fluorescence of the protein in the crystals. The position of the maximum and the bandwidth of the steady-state emission spectrum of the crystals (331 ± 1 and 43 ± 1 nm, respectively) are equal to those obtained in aqueous buffer for the orosomucoid–progesterone complex (330 ± 1 and 43 ± 1 nm) (ex, 295 nm). Thus, the fluorescence of the crystals occurs from the Trp residues buried in the protein core. The red-edge excitation spectra studies indicate that the Trp residues are surrounded by microenvironments that display motions, a result identical to that observed in solution. Thus, the crystallization process does not modify the structure or the dynamics of orosomucoid core. The fluorescence intensities depend on the angular orientation of the crystals with respect to the polarization of the incident beam. The general feature of this dependence is identical at the three excitation wavelengths used (295, 300, and 305 nm). Our results confirm the fact that the local structure and dynamics are the key for any interpretation of tryptophan fluorescence parameters of orosomucoid. 相似文献
112.
色氨酸和酪氨酸的三维荧光光谱特征参量提取 总被引:3,自引:0,他引:3
氨基酸是维持生命活动的重要物质,而色氨酸和酪氨酸又是天然氨基酸中重要的发光组分,应用荧光光谱法对其进行测量和分辨具有重要的意义。文章用美国Pekin-Elmer LS55型荧光分光光度计,对色氨酸和酪氨酸的三维荧光光谱进行了测量。将测量的数据用激发-发射-荧光强度的三维坐标表示,得到三维荧光谱图,但色氨酸和酪氨酸存在共性峰,通过波峰位置简单地来辨别两种混叠的物质很有难度。以数理统计概念为基础,提取该三维荧光光谱的特征参数,得到两种物质荧光光谱中最相关的信息,可以解决两种物质光谱混叠的分辨问题。结果表明,色氨酸和酪氨酸的三维荧光光谱平均值、标准差、原点矩、混合中心矩等参数差值百分比分别为330.37%, 102.86%, 329.16%, 329.63%,区别较大;而边际分布、相关系数值差值百分比仅为10.61%和2.40%。因而平均值、标准差、原点矩、混合中心矩可作为敏感特征参数,用其分辨谱图混叠的色氨酸和酪氨酸是可行的。这种“数学预提取”的三维光谱分析法可以找出组分之间的敏感特征参量,能够取代传统的三维荧光光谱分析法。 相似文献
113.
二阶导数荧光分光光度法同时测定色氨酸和酷氨酸 总被引:4,自引:2,他引:2
本文描述了用二阶导数荧光光度法同时测定色氨酸和酪氨酸。在 p H 7.4的条件下 ,用 2 2 1nm作为激发波长 ,记录色氨酸和酪氨酸的发射光谱 ,并进行二阶导数处理。色氨酸在 318nm处 ,酪氨酸在 2 83nm处 ,二阶导数峰高与浓度成线性关系。色氨酸工作曲线的线性回归方程为 c =0 .0 0 0 7H - 0 .0 0 4,r =0 .996 4,线性范围为 0 .0 0 4到 0 .2 0 0μg . m L- 1 。酪氨酸工作曲线的线性回归方程为 c =0 .0 0 12 H -0 .0 0 40 ,r=0 .9971。线性范围为 0 .0 0 2到 0 .2 5 0 μg· m L- 1 。实验了 p H、温度和干扰离子对测定的影响 ,测定了苹果中的色氨酸和酪氨酸的含量 ,回收率分别为 (92 .0~ 10 4.0 ) %和 (98.70~ 10 2 .0 ) % ,相对标准偏差分别为 3.5 %和 2 .8%。 相似文献
114.
LIANGFang SUNHong ZHAOCheng-guang CUITing HONGShui-sheng CHENJia LIULan-ying 《高等学校化学研究》2003,19(3):317-319
The chemical modification of human plasminogen (HPg) was studied with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), N-acetylimidazole (NAI), 1,2-cyclohexanedione (CHD), chloramine T(Ch-T)and N-bromosuccinimide (NBS) as modifying reagents at its carboxyl group, tyrosine, arginine, methionine and tryptophan residues, respectively. The results indicate that tyrosine and arginine residues are not essential for HPg activity, while carboxyl groups, methionine and tryptophan residues are important for the activi-ty of HPg. The Keech and Farrant‘s kinetic analysis reveals that one tryptophan residue, one methionine residue and two carboxyl groups are essential for HPg activity. 相似文献
115.
HE Xiao-hong WU Min LI Shan-yu CHU Yu-zhuo CHEN Jia LIU Lan-ying 《高等学校化学研究》2005,21(5):562-565
The amino acid composition of the superoxide dismutase(SOD) from camellia pollen was measured and the tryptophan(Trp) residues were modified by using N-bromosuccinimide(NBS). The results show that there are 21 Trp residues in an SOD molecule and seven of which are located on the surface of the enzyme. By researching the fluorescence spectra of the native SOD and the modified SOD, we have found that the emission wavelength of Trp is at 335 nm and the fluorescence intensity will decrease when the enzyme is modified. The results also show that potassium iodide(KI) can significantly quench the fluorescence of the native SOD, but it has a less pronounced effect on the modified enzyme. Glycerin as a surface activation reagent can stabilize the fluorescence of the modified enzyme. 相似文献
116.
117.
Sunlight held the key to the origin of life on Earth. The earliest life forms, cyanobacteria, captured the sunlight to generate energy through photosynthesis. Life on Earth evolved in accordance with the circadian rhythms tied to sensitivity to sunlight patterns. A unique feature of cyanobacterial photosynthetic proteins and circadian rhythms' molecules, and later of nearly all photon-sensing molecules throughout evolution, is that the aromatic amino acid tryptophan (Trp) resides at the center of light-harvesting active sites. In this perspective, I review the literature and integrate evidence from different scientific fields to explore the role Trp plays in photon-sensing capabilities of living organisms through its resonance delocalization of π-electrons. The observations presented here are the product of apparently unrelated phenomena throughout evolution, but nevertheless share consistent patterns of photon-sensing by Trp-containing and Trp-derived molecules. I posit the unique capacity to transfer electrons during photosynthesis in the earliest life forms is conferred to Trp due to its aromaticity. I propose this ability evolved to assume more complex functions, serving as a host for mechanisms underlying mental aptitudes – a concept which provides a theoretical basis for defining the neural correlates of consciousness. The argument made here is that Trp aromaticity may have allowed for the inception of the mechanistic building blocks used to fabricate complexity in higher forms of life. More specifically, Trp aromatic non-locality may have acted as a catalyst for the emergence of consciousness by instigating long-range synchronization and stabilizing the large-scale coherence of neural networks, which mediate functional brain activity. The concepts proposed in this perspective provide a conceptual foundation that invites further interdisciplinary dialogue aimed at examining and defining the role of aromaticity (beyond Trp) in the emergence of life and consciousness. 相似文献
118.
119.
<正>In this study,we have fabricated molecularly imprinted polypyrrole(PPy) packed electrode columns and investigated their effects on separation of tryptophan(Trp) enantiomers by using potential control.The results indicate that the imprinted PPy electrode columns could efficiently enhance the L-Trp uptake and separate Trp enantiomers effectively,implying the great potential for the enantioselective recognition of other amino acids enantiomers. 相似文献
120.
Determination of cysteamine and tryptophan is described by electrochemical methods using p‐aminophenol‐multiwall carbon nanotubes paste electrode. Cysteamine and tryptophan in mixture can each be measured independently from each other with a potential difference of 600 mV. The results showed that the electrocatalytic currents increased linearly with cysteamine and tryptophan concentrations over the ranges 0.5–300 µmol L?1 and 10.0–650 µmol L?1, respectively. The detection limits for cysteamine and tryptophan are found to be 0.14 and 5.9 µmol L?1, respectively. The proposed method is successfully employed for the determination of cysteamine in both capsule and urine samples. 相似文献