全文获取类型
收费全文 | 1232篇 |
免费 | 57篇 |
国内免费 | 1篇 |
专业分类
1290篇 |
出版年
2022年 | 11篇 |
2021年 | 15篇 |
2020年 | 9篇 |
2019年 | 17篇 |
2018年 | 18篇 |
2017年 | 13篇 |
2016年 | 23篇 |
2015年 | 54篇 |
2014年 | 60篇 |
2013年 | 170篇 |
2012年 | 91篇 |
2011年 | 77篇 |
2010年 | 68篇 |
2009年 | 37篇 |
2008年 | 79篇 |
2007年 | 77篇 |
2006年 | 55篇 |
2005年 | 59篇 |
2004年 | 60篇 |
2003年 | 57篇 |
2002年 | 46篇 |
2001年 | 7篇 |
2000年 | 11篇 |
1999年 | 10篇 |
1998年 | 7篇 |
1997年 | 10篇 |
1996年 | 3篇 |
1995年 | 9篇 |
1994年 | 9篇 |
1993年 | 9篇 |
1992年 | 10篇 |
1991年 | 9篇 |
1990年 | 11篇 |
1989年 | 9篇 |
1988年 | 5篇 |
1987年 | 8篇 |
1986年 | 4篇 |
1985年 | 5篇 |
1984年 | 6篇 |
1983年 | 4篇 |
1982年 | 5篇 |
1981年 | 9篇 |
1979年 | 4篇 |
1978年 | 2篇 |
1977年 | 2篇 |
1976年 | 5篇 |
1974年 | 9篇 |
1973年 | 3篇 |
1972年 | 2篇 |
1970年 | 2篇 |
排序方式: 共有1290条查询结果,搜索用时 15 毫秒
991.
Olivier Serve Yukiko Kamiya Aya Maeno Michiko Nakano Chiho Murakami Yoshiki Yamaguchi Takushi Harada Maho Yagi-Utsumi Kenji Inaba Osamu Asami Toshihiko Oka Koichi Kato 《Journal of molecular biology》2010,396(2):361-2193
Protein disulfide isomerase (PDI) is a major protein in the endoplasmic reticulum, operating as an essential folding catalyst and molecular chaperone for disulfide-containing proteins by catalyzing the formation, rearrangement, and breakage of their disulfide bridges. This enzyme has a modular structure with four thioredoxin-like domains, a, b, b′, and a′, along with a C-terminal extension. The homologous a and a′ domains contain one cysteine pair in their active site directly involved in thiol-disulfide exchange reactions, while the b′ domain putatively provides a primary binding site for unstructured regions of the substrate polypeptides. Here, we report a redox-dependent intramolecular rearrangement of the b′ and a′ domains of PDI from Humicola insolens, a thermophilic fungus, elucidated by combined use of nuclear magnetic resonance (NMR) and small-angle X-ray scattering (SAXS) methods. Our NMR data showed that the substrates bound to a hydrophobic surface spanning these two domains, which became more exposed to the solvent upon oxidation of the active site of the a′ domain. The hydrogen-deuterium exchange and relaxation data indicated that the redox state of the a′ domain influences the dynamic properties of the b′ domain. Moreover, the SAXS profiles revealed that oxidation of the a′ active site causes segregation of the two domains. On the basis of these data, we propose a mechanistic model of PDI action; the a′ domain transfers its own disulfide bond into the unfolded protein accommodated on the hydrophobic surface of the substrate-binding region, which consequently changes into a “closed” form releasing the oxidized substrate. 相似文献
992.
Akazawa S Karino T Yoshida N Katsuragi T Tani Y 《Applied and environmental microbiology》2004,70(10):5882-5890
Three active fractions of fructosyl-amino acid oxidase (FAOD-Ao1, -Ao2a, and -Ao2b) were isolated from Aspergillus oryzae strain RIB40. N-terminal and internal amino acid sequences of FAOD-Ao2a corresponded to those of FAOD-Ao2b, suggesting that these two isozymes were derived from the same protein. FAOD-Ao1 and -Ao2 were different in substrate specificity and subunit assembly; FAOD-Ao2 was active toward N(epsilon)-fructosyl N(alpha)-Z-lysine and fructosyl valine (Fru-Val), whereas FAOD-Ao1 was not active toward Fru-Val. The genes encoding the FAOD isozymes (i.e., FAOAo1 and FAOAo2) were cloned by PCR with an FAOD-specific primer set. The deduced amino acid sequences revealed that FAOD-Ao1 was 50% identical to FAOD-Ao2, and each isozyme had a peroxisome-targeting signal-1, indicating their localization in peroxisomes. The genes was expressed in Escherichia coli and rFaoAo2 showed the same characteristics as FAOD-Ao2, whereas rFaoAo1 was not active. FAOAo2 disruptant was obtained by using ptrA as a selective marker. Wild-type strain grew on the medium containing Fru-Val as the sole carbon and nitrogen sources, but strain Delta faoAo2 did not grow. Addition of glucose or (NH(4))(2)SO(4) to the Fru-Val medium did not affect the assimilation of Fru-Val by wild-type, indicating glucose and ammonium repressions did not occur in the expression of the FAOAo2 gene. Furthermore, conidia of the wild-type strain did not germinate on the medium containing Fru-Val and NaNO(2) as the sole carbon and nitrogen sources, respectively, suggesting that Fru-Val may also repress gene expression of nitrite reductase. These results indicated that FAOD is needed for utilization of fructosyl-amino acids as nitrogen sources in A. oryzae. 相似文献
993.
The Rho-mDia1 pathway regulates cell polarity and focal adhesion turnover in migrating cells through mobilizing Apc and c-Src 下载免费PDF全文
Yamana N Arakawa Y Nishino T Kurokawa K Tanji M Itoh RE Monypenny J Ishizaki T Bito H Nozaki K Hashimoto N Matsuda M Narumiya S 《Molecular and cellular biology》2006,26(18):6844-6858
Directed cell migration requires cell polarization and adhesion turnover, in which the actin cytoskeleton and microtubules work critically. The Rho GTPases induce specific types of actin cytoskeleton and regulate microtubule dynamics. In migrating cells, Cdc42 regulates cell polarity and Rac works in membrane protrusion. However, the role of Rho in migration is little known. Rho acts on two major effectors, ROCK and mDia1, among which mDia1 produces straight actin filaments and aligns microtubules. Here we depleted mDia1 by RNA interference and found that mDia1 depletion impaired directed migration of rat C6 glioma cells by inhibiting both cell polarization and adhesion turnover. Apc and active Cdc42, which work together for cell polarization, localized in the front of migrating cells, while active c-Src, which regulates adhesion turnover, localized in focal adhesions. mDia1 depletion impaired localization of these molecules at their respective sites. Conversely, expression of active mDia1 facilitated microtubule-dependent accumulation of Apc and active Cdc42 in the polar ends of the cells and actin-dependent recruitment of c-Src in adhesions. Thus, the Rho-mDia1 pathway regulates polarization and adhesion turnover by aligning microtubules and actin filaments and delivering Apc/Cdc42 and c-Src to their respective sites of action. 相似文献
994.
Yoshiki Koriyama Kayo Sugitani Kazuhiro Ogai Satoru Kato 《Journal of neurochemistry》2014,130(5):707-719
Retinal degenerative diseases (RDs) are a group of inherited diseases characterized by the loss of photoreceptor cells. Selective photoreceptor loss can be induced in mice by an intraperitoneal injection of N‐methyl‐N‐nitrosourea (MNU) and, because of its selectivity, this model is widely used to study the mechanism of RDs. Although it is known that calcium‐calpain activation and lipid peroxidation are involved in the initiation of cell death, the precise mechanisms of this process remain unknown. Heat shock protein 70 (HSP70) has been shown to function as a chaperone molecule to protect cells against environmental and physiological stresses. In this study, we investigated the role of HSP70 on photoreceptor cell death in mice. HSP70 induction by valproic acid, a histone deacetylase inhibitor, attenuated the photoreceptor cell death by MNU through inhibition of apoptotic caspase signals. Furthermore, HSP70 itself was rapidly and calpain‐dependently cleaved after MNU treatment. Therefore, HSP70 induction by valproic acid was dually effective against MNU‐induced photoreceptor cell loss as a result of its anti‐apoptotic actions and its ability to prevent HSP70 degradation. These findings might help lead us to a better understanding of the pathogenic mechanism of RDs.
995.
996.
Yamasaki Y 《Phytochemistry》2003,64(5):935-939
Beta-amylase (EC 3.2.1.2) was isolated from germinating millet (Panicum miliaceum L.) seeds by a procedure that included ammonium sulfate fractionation, chromatography on DEAE-cellulofine and CM-cellulofine, and preparative isoelectric focusing. The enzyme was homogeneous by SDS-PAGE. The M(r) of the enzyme was estimated to be 58,000 based on its mobility on SDS-PAGE and gel filtration with TSKgel G4000SW(XL), which showed that it is composed of a single unit. The isoelectric point of the enzyme was 4.62. The enzyme hydrolyzed malto-oligosaccharides more readily as their degree of polymerization increased, this being strongest for malto-oligosaccharides larger than 13 glucose residues and very weakly for maltotriose. Amylose, amylopectin and soluble starch were the most suitable substrates for the enzyme. While the enzyme showed some activity against native starch by itself, starch digestion was accelerated 2.5-fold using alpha-amylase, pullulanase and alpha-glucosidase. This enzyme appears to be very important for the germination of millet seeds. 相似文献
997.
Yoshiki Nakamura Satoshi Simpo Minyeong Lee Takashi Oikawa Taeko Yoshii Koji Noda Yosuke Kuwahara Kenzo Kawasaki 《Biotechnic & histochemistry》2000,75(1):1-6
We observed the histology and tetracycline (TC) labeling in a single frontal section of alveolar bone of upper first molars of adolescent rats. A single injection of TC was administered intraperitonealy in adolescent rats. After three weeks, the upper jaws were immersed rapidly in liquid nitrogen and sectioned. Five micrometer unfixed, undecalcified frozen sections were cut and observed by light and fluorescence microscopy. Frontal sections of the upper first molar area revealed that the structural relationships among the roots, the periodontal ligament and the alveolar bone, and also between the cervical enamel and the attachment epithelium were well preserved. The TC labeling lines in the sections were very clear and distinguished new bone from old bone. The brightness of the lines differed among regions. An analysis of the brightness in the same section suggested a difference in the bone forming activity at the time of injection. 相似文献
998.
999.
Yoshiki Yamaguchi Masami Masuda Hiroaki Sasakawa Takashi Nonaka Shin-ichi Hisanaga Masato Hasegawa 《Journal of molecular biology》2010,395(3):445-456
α-Synuclein is a major component of filamentous inclusions that are histological hallmarks of Parkinson's disease and other α-synucleinopathies. Previous analyses have revealed that several polyphenols inhibit α-synuclein assembly with low micromolar IC50 values, and that SDS-stable, noncytotoxic soluble α-synuclein oligomers are formed in their presence. Structural elucidation of inhibitor-bound α-synuclein oligomers is obviously required for the better understanding of the inhibitory mechanism. In order to characterize inhibitor-bound α-synucleins in detail, we have prepared α-synuclein dimers in the presence of polyphenol inhibitors, exifone, gossypetin, and dopamine, and purified the products. Peptide mapping and mass spectrometric analysis revealed that exifone-treated α-synuclein monomer and dimer were oxidized at all four methionine residues of α-synuclein. Immunoblot analysis and redox-cycling staining of endoproteinase Asp-N-digested products showed that the N-terminal region (1-60) is involved in the dimerization and exifone binding of α-synuclein. Ultra-high-field NMR analysis of inhibitor-bound α-synuclein dimers showed that the signals derived from the N-terminal region of α-synuclein exhibited line broadening, confirming that the N-terminal region is involved in inhibitor-induced dimerization. The C-terminal portion still predominantly exhibited the random-coil character observed in monomeric α-synuclein. We propose that the N-terminal region of α-synuclein plays a key role in the formation of α-synuclein assemblies. 相似文献
1000.
Okamoto K Inaba M Furumitsu Y Ban A Mori N Yukioka K Imanishi Y Nishizawa Y 《Life sciences》2010,87(23-26):686-691