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41.
The Role of the Membrane-spanning Domain Sequence in Glycoprotein-mediated Membrane Fusion 总被引:1,自引:1,他引:1
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The role of glycoprotein membrane-spanning domains in the process of membrane fusion is poorly understood. It has been demonstrated that replacing all or part of the membrane-spanning domain of a viral fusion protein with sequences that encode signals for glycosylphosphatidylinositol linkage attachment abrogates membrane fusion activity. It has been suggested, however, that the actual amino acid sequence of the membrane-spanning domain is not critical for the activity of viral fusion proteins. We have examined the function of Moloney murine leukemia virus envelope proteins with substitutions in the membrane-spanning domain. Envelope proteins bearing substitutions for proline 617 are processed and incorporated into virus particles normally and bind to the viral receptor. However, they possess greatly reduced or undetectable capacities for the promotion of membrane fusion and infectious virus particle formation. Our results imply a direct role for the residues in the membrane-spanning domain of the murine leukemia virus envelope protein in membrane fusion and its regulation. They also support the thesis that membrane-spanning domains possess a sequence-dependent function in other protein-mediated membrane fusion events. 相似文献
42.
The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis. 总被引:21,自引:2,他引:19
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V Sudakin D Ganoth A Dahan H Heller J Hershko F C Luca J V Ruderman A Hershko 《Molecular biology of the cell》1995,6(2):185-197
The ubiquitin-mediated degradation of mitotic cyclins is required for cells to exit from mitosis. Previous work with cell-free systems has revealed four components required for cyclin-ubiquitin ligation and proteolysis: a nonspecific ubiquitin-activating enzyme E1, a soluble fraction containing a ubiquitin carrier protein activity called E2-C, a crude particulate fraction containing a ubiquitin ligase (E3) activity that is activated during M-phase, and a constitutively active 26S proteasome that degrades ubiquitinated proteins. Here, we identify a novel approximately 1500-kDa complex, termed the cyclosome, which contains a cyclin-selective ubiquitin ligase activity, E3-C. E3-C is present but inactive during interphase; it can be activated in vitro by the addition of cdc2, enabling the transfer of ubiquitin from E2-C to cyclin. The kinetics of E3-C activation suggest the existence of one or more intermediates between cdc2 and E3-C. Cyclosome-associated E3-C acts on both cyclin A and B, and requires the presence of wild-type N-terminal destruction box motifs in each cyclin. Ubiquitinated cyclins are then rapidly recognized and degraded by the proteasome. These results identify the cyclosome-associated E3-C as the component of the cyclin destruction machinery whose activity is ultimately regulated by cdc2 and, as such, the element directly responsible for setting mitotic cyclin levels during early embryonic cell cycles. 相似文献
43.
R. Weiss Dominique Mandon Thomas Wolter Alfred X. Trautwein Markus Müther Eckhard Bill Avram Gold Karupiah Jayaraj James Terner 《Journal of biological inorganic chemistry》1996,1(4):377-383
A comparison of the exchange interactions arising in the peroxidase and catalase Compound I intermediates and their iron(IV)-oxo
porphyrin π-cation radical models, both of which are two oxidizing equivalents above the ferric state, suggests that in the
models the oxidizing equivalent is localized on the porphyrin ring, while in the proteins it is partly delocalized onto the
proximal ligand. Thus, the proximal axial ligand of iron participates indirectly in the oxidation reactions catalyzed by the
enzymes. Possible roles of the axial ligand in the catalytic mechanism of these heme-based enzymes are discussed.
Received and accepted: 7 May 1996 相似文献
44.
45.
Conversion of Leumorphin (Dynorphin B-29) to Dynorphin B and Dynorphin B-14 by Thiol Protease Activity 总被引:2,自引:2,他引:0
Dynorphin B (rimorphin) is formed from leumorphin (dynorphin B-29) by the action of a thiol protease from rat brain membranes, in a single step. This represents a "single-arginine cleavage" between threonine-13 and arginine-14 of the substrate. We have observed that in addition to dynorphin B, dynorphin B-14 is formed from dynorphin B-29. Among the various protease inhibitors tested, none except p-chloromercuribenzensulfonic acid inhibited the formation of the two products. Both temperature and pH had similar effects on the formation of dynorphin B-14 and dynorphin B. The inhibitory potencies of adrenocorticotropic hormone, peptide E, and dynorphin A were virtually identical for the formation of the two products. These results suggest that the same enzyme may be responsible for the formation of dynorphin B-14 and dynorphin B. 相似文献
46.
Influence of protease inhibitors and energy metabolism on intracellular protein breakdown in starving Escherichia coli 总被引:6,自引:0,他引:6
Y Shechter D Rafaeli-Eshkol A Hershko 《Biochemical and biophysical research communications》1973,54(4):1518-1524
It has been reported that certain inhibitors of serine proteases block intracellular protein breakdown in subjected to nutritional deprivation. We show here that the protease inhibitors p-toluene sulfonyl fluoride and pentamidine isethionate inhibit protein breakdown in deprived of glucose, but not in bacteria starved for inorganic phosphate or ammonia. Furthermore, we find that the protease inhibitors cause a drastic decline in cellular ATP levels when glucose is omitted from the incubation medium. It is concluded that these protease inhibitors influence protein breakdown by interfering with cellular energy production, rather than by interacting with a specific serine protease. 相似文献
47.
Avram Hershko 《Biochimica et Biophysica Acta (BBA)/General Subjects》1977,498(1):83-90
Corticosteroi-induced tyrosine aminotransferase (EC 2.6.1.5) from cultured hepatoma cells was separated by carboxymethyl-Sephadex chromatography into three molecular forms resembling those described previously in the rat liver. Enzyme forms were isolated and used as purified substrates to examine their in vitro interconversion by various subcellular fractions. Isolated form III was converted to forms II and I, and isolated form II was converted to form I by the coarse particulate fraction sedimenting at 1000 × g. This activity was inhibited by the serine enzyme inhibitor phenylmethane sulfonyl fluoride or by raising the pH to 8.7. Conversion of enzyme forms in vitro in the opposite direction (I → II → III) could not be detected. The distribution of enzyme forms in vivo was examined by the use of experimental conditions that prevent their in vitro interconversion during cell extraction. Tyrosine aminotransferase extracted from cells subjected to various treatments that affect the rates of enzyme synthesis or degradation existed always predominantly as form III. It appears, therefore, that multiple forms of tyrosine aminotransferase are not related to the turnover of this enzyme in vivo. 相似文献
48.
Two L-methionine-enriched mutants, SN-78 and SE-57, were isolated in a sulphur-deficient medium from the methylotrophic yeast Candida boidinii ICCF26. No significant differences were detected between the L-cysteine pools of the mutants and the wild-type. In mutant strain SE-57, S-adenosylmethionine and ergosterol levels were higher than in the wild-type strain, while in the other mutant, SN-78, the levels were lower. The evidence presented would suggest that, in both the mutants and the wild-type used in this study, S-adenosylmethionine was of key importance for the accumulation of L-methionine and ergosterol. 相似文献
49.
Characterization of the heat-stable polypeptide of the ATP-dependent proteolytic system from reticulocytes 总被引:10,自引:0,他引:10
A Ciechanover S Elias H Heller S Ferber A Hershko 《The Journal of biological chemistry》1980,255(16):7525-7528
A heat-stable polypeptide from rabbit reticulocytes has been previously shown to be required for ATP-dependent protein breakdown (Ciechanover, A., Hod, Y., and Hershko, A. (1978) Biochem. Biophys. Res Commun. 81, 1100-1105) and to form covalent conjugates with proteins in an ATP-requiring reaction (Ciechanover, A., Heller, H., Elias, S., Haas, A.L., and Hershko, A. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 1365-1368). We now describe its purification, characterization, and tissue distribution. Its presence in erythrocytes at high level makes this a possible preferred source in the future. 相似文献
50.