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排序方式: 共有166条查询结果,搜索用时 31 毫秒
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124.
G. Flood A. J. Rowe D. R. Critchley W. B. Gratzer 《European biophysics journal : EBJ》1997,25(5-6):431-435
Protein constructs consisting of repeats 1–4, repeats 1–3 and repeats 2–4 of the rod domain of chicken α-actinin were expressed as fusion proteins in Escherichia coli. Based on the evidence of circular dichroism spectra and cooperative thermal unfolding profiles both truncated rod fragments
were judged to have assumed the native structural fold. The thermal stabilities were in both cases significantly lower than
that of the intact rod (repeats 1–4). Analyses by sedimentation equilibrium and velocity provided further evidence to show
that fragment 1–4 is entirely dimeric in the concentration range of these experiments, resembling therefore the rod domain
isolated by proteolytic digestion of native α-actinin. Fragment 2–4, and probably also 1–3, show concentration-dependent association, with dissociation constants, estimated
by sedimentation equilibrium, in the 1–10 μM range. Thus, in confirmation of earlier work, all four repeats are re-quired
to generate a maximally stable anti-parallel dimer (Kd∼10 pM), suggesting the presence of binding sites in all of them to allow for aligned pairing.
Accepted: 7 November 1996 相似文献
125.
Schmid AW Condemi E Tuchscherer G Chiappe D Mutter M Vogel H Moniatte M Tsybin YO 《The Journal of biological chemistry》2011,286(14):12172-12188
Tissue transglutaminase (TGase) has been implicated in a number of cellular processes and disease states, where the enzymatic actions of TGase may serve in both, cell survival and apoptosis. To date, the precise functional properties of TGase in cell survival or cell death mechanisms still remain elusive. TGase-mediated cross-linking has been reported to account for the formation of insoluble lesions in conformational diseases. We report here that TGase induces intramolecular cross-linking of β-amyloid peptide (Aβ), resulting in structural changes of monomeric Aβ. Using high resolution mass spectrometry (MS) of cross-linked Aβ peptides, we observed a shift in mass, which is, presumably associated with the loss of NH3 due to enzymatic transamidation activity and hence intramolecular peptide cross-linking. We have observed that a large population of Aβ monomers contained an 0.984 Da increase in mass at a glutamine residue, indicating that glutamine 15 serves as an indispensable substrate in TGase-mediated deamidation to glutamate 15. We provide strong analytical evidence on TGase-mediated Aβ peptide dimerization, through covalent intermolecular cross-linking and hence the formation of Aβ1-40 dimers. Our in depth analyses indicate that TGase-induced post-translational modifications of Aβ peptide may serve as an important seed for aggregation. 相似文献
126.
Marquer C Fruchart-Gaillard C Letellier G Marcon E Mourier G Zinn-Justin S Ménez A Servent D Gilquin B 《The Journal of biological chemistry》2011,286(36):31661-31675
The snake toxin MT7 is a potent and specific allosteric modulator of the human M1 muscarinic receptor (hM1). We previously characterized by mutagenesis experiments the functional determinants of the MT7-hM1 receptor interaction (Fruchart-Gaillard, C., Mourier, G., Marquer, C., Stura, E., Birdsall, N. J., and Servent, D. (2008) Mol. Pharmacol. 74, 1554–1563) and more recently collected evidence indicating that MT7 may bind to a dimeric form of hM1 (Marquer, C., Fruchart-Gaillard, C., Mourier, G., Grandjean, O., Girard, E., le Maire, M., Brown, S., and Servent, D. (2010) Biol. Cell 102, 409–420). To structurally characterize the MT7-hM1 complex, we adopted a strategy combining double mutant cycle experiments and molecular modeling calculations. First, thirty-three ligand-receptor proximities were identified from the analysis of sixty-one double mutant binding affinities. Several toxin residues that are more than 25 Å apart still contact the same residues on the receptor. As a consequence, attempts to satisfy all the restraints by docking the toxin onto a single receptor failed. The toxin was then positioned onto two receptors during five independent flexible docking simulations. The different possible ligand and receptor extracellular loop conformations were described by performing simulations in explicit solvent. All the docking calculations converged to the same conformation of the MT7-hM1 dimer complex, satisfying the experimental restraints and in which (i) the toxin interacts with the extracellular side of the receptor, (ii) the tips of MT7 loops II and III contact one hM1 protomer, whereas the tip of loop I binds to the other protomer, and (iii) the hM1 dimeric interface involves the transmembrane helices TM6 and TM7. These results structurally support the high affinity and selectivity of the MT7-hM1 interaction and highlight the atypical mode of interaction of this allosteric ligand on its G protein-coupled receptor target. 相似文献
127.
SM Jung M Moroi K Soejima T Nakagaki Y Miura MC Berndt EE Gardiner JM Howes N Pugh D Bihan SP Watson RW Farndale 《The Journal of biological chemistry》2012,287(35):30000-30013
The platelet collagen receptor glycoprotein VI (GPVI) has been suggested to function as a dimer, with increased affinity for collagen. Dissociation constants (K(d)) obtained by measuring recombinant GPVI binding to collagenous substrates showed that GPVI dimers bind with high affinity to tandem GPO (Gly-Pro-Hyp) sequences in collagen, whereas the markedly lower affinity of the monomer for all substrates implies that it is not the collagen-binding form of GPVI. Dimer binding required a high density of immobilized triple-helical (GPO)(10)-containing peptide, suggesting that the dimer binds multiple, discrete peptide helices. Differential inhibition of dimer binding by dimer-specific antibodies, m-Fab-F and 204-11 Fab, suggests that m-Fab-F binds at the collagen-binding site of the dimer, and 204-11 Fab binds to a discrete site. Flow cytometric quantitation indicated that GPVI dimers account for ~29% of total GPVI in resting platelets, whereas activation by either collagen-related peptide or thrombin increases the number of dimers to ~39 and ~44%, respectively. m-Fab-F inhibits both GPVI-dependent static platelet adhesion to collagen and thrombus formation on collagen under low and high shear, indicating that pre-existing dimeric GPVI is required for the initial interaction with collagen because affinity of the monomer is too low to support binding and that interaction through the dimer is essential for platelet activation. These GPVI dimers in resting circulating platelets will enable them to bind injury-exposed subendothelial collagen to initiate platelet activation. The GPVI-specific agonist collagen-related peptide or thrombin further increases the number of dimers, thereby providing a feedback mechanism for reinforcing binding to collagen and platelet activation. 相似文献
128.
Harper FM Addison JA Hart MW 《Evolution; international journal of organic evolution》2007,61(10):2410-2418
Phylogeographic studies designed to estimate rates and patterns of genetic differentiation within species often reveal unexpected and graphically striking cases of allele or haplotype sharing between species (introgression) via hybridization and backcrossing. Does introgression between species significantly influence population genetic structure relative to more conventional sources of differentiation (drift) and similarity (dispersal) among populations within species? Here we use mtDNA sequences from four species in two genera of sea urchins and sea stars to quantify the relative magnitude of gene flow across oceans and across species boundaries in the context of the trans-Arctic interchange of marine organisms between the Pacific and Atlantic oceans. In spite of the much smaller distances between sympatric congeners, rates of gene flow between sympatric species via heterospecific gamete interactions were small and significantly lower than gene flow across oceans via dispersal of planktonic larvae. We conclude that, in these cases at least, larvae are more effective than gametes as vectors of gene flow. 相似文献
129.
Zhaosheng Qian 《Inorganica chimica acta》2010,363(13):3627-4189
A dissociative (D) and a solvent-assisted dissociative interchange (Id) water-exchange pathways for magnesium(II) in aqueous solution were simulated with density functional theory calculations. The D mechanism of includes a five-coordinated intermediate, while the Id water-exchange pathway of proceeds with the assistance of a solvent water molecule, which supports the experimental assignment of the reaction mechanism. The intrinsic activation volume was used to differentiate between Id and Ia mechanisms despite of the exclusion of the contribution of transmission coefficient. The calculated intrinsic activation volume for the Id mechanism is consistent with the experimental data, and is closer to the experimental data than that for D mechanism. The Id mechanism is suggested as the dominate water-exchange pathway of depending on the intrinsic activation volume with the assistance of the activation entropy. The calculations also showed that the influences of the explicit and bulk waters on energy barriers for D and Id mechanisms are obviously different. 相似文献
130.
John Calambokidis Gretchen H. Steiger Joseph R. Evenson Kiirsten R. Flynn Kenneth C. Balcomb Diane E. Claridge Prentice Bloedel Janice M. Straley C. Scott Baker Olga VON Ziegesar Marilyn E. Dahlheim Janice M. Waite James D. Darling Graeme Ellis Gregory A. Green 《Marine Mammal Science》1996,12(2):215-226
Humpback whales feed in several high-latitude areas of the North Pacific. We examined the interchange of humpback whales between one of these areas, off California, and those in other feeding grounds in the eastern North Pacific:. Fluke photographs of 597 humpback whales identified off California between 1986 and 1992 were compared with those off Oregon and Washington (29); British Columbia (81); southeastern Alaska (343); Prince William Sound, Alaska (141); Kodiak Island, Alaska (104); Shumagin Islands, Alaska (22); and in the Bering Sea (7). A high degree of interchange, both inter-and intrayear, was found among humpback whales seen off California, Oregon, and Washington., A low rate of interchange was found between British Columbia and California.: two whales seen near the British Columbia/Washington border were photographed off California in a different year, No interchange was found between California and the three feeding areas in Alaska. Humpback whales off California, Oregon, and Washington form a single intermixing feeding aggregation with only limited interchange with areas farther north. These findings are consistent with photographic identification studies in the North Atlantic and with genetic studies in both the North Atlantic and North Pacific. 相似文献