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91.
CMA Iwegbue OV Beecroft JE Ogala FE Egobueze GO Tesi GE Nwajei 《African Journal of Aquatic Science》2018,43(3):241-253
The concentrations of nine metals were measured by atomic absorption spectrophotometry in surface sediments of three coastal creeks, namely, the Ifie, Egbokodo and Ubeji creeks, in the Niger Delta of Nigeria, from August 2012 to January 2013. The aim of the study was to provide information on the spatial and seasonal distribution patterns, degree of contamination, and ecological risks of metals in these sediments. The mean concentrations of the nine metals in these creek sediments ranged from 0.30 to 3.20?mg kg?1 Cd; 10.7 to 24.7?mg kg?1 Pb, 125 to 466?mg kg?1 Cr; 3.1.10 to 14.9?mg kg?1 Cu; 4.7 to 14.3?mg kg?1 Co; 61.1 to 115?mg kg?1 Ni; 106 to 183?mg kg?1 Mn; 52.0 to 170?mg kg?1 Zn and 5 469 to 20 639?mg kg?1 Fe. In general, the metal concentrations were higher in the dry season than the wet season, except for Cr. The concentrations of Cd, Cr, Ni and Zn were above their regulatory control limits in sediment as specified by the Nigerian Regulatory Authority and Cd was identified as the main ecological risk factor. The enrichment factors for the studied metals followed the order: Cd > Cr > Ni > Zn > Pb > Co > Mn > Cu. The average multiple pollution index values indicated that these sediments were severely polluted with significant inputs from Cd, Ni and Cr. 相似文献
92.
Mutations of fumarase that distinguish between the active site and a nearby dicarboxylic acid binding site. 总被引:3,自引:1,他引:2
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T. Weaver M. Lees L. Banaszak 《Protein science : a publication of the Protein Society》1997,6(4):834-842
Two mutant forms of fumarase C from E. coli have been made using PCR and recombinant DNA. The recombinant form of the protein included a histidine arm on the C-terminal facilitating purification. Based on earlier studies, two different carboxylic acid binding sites, labeled A- and B-, were observed in crystal structures of the wild type and inhibited forms of the enzyme. A histidine at each of the sites was mutated to an asparagine. H188N at the A-site resulted in a large decrease in specific activity, while the H129N mutation at the B-site had essentially no effect. From the results, we conclude that the A-site is indeed the active site, and a dual role for H188 as a potential catalytic base is proposed. Crystal structures of the two mutant proteins produced some unexpected results. Both mutations reduced the affinity for the carboxylic acids at their respective sites. The H129N mutant should be particularly useful in future kinetic studies because it sterically blocks the B-site with the carboxyamide of asparagine assuming the position of the ligand's carboxylate. In the H188N mutation at the active site, the new asparagine side chain still interacts with an active site water that appears to have moved slightly as a result of the mutation. 相似文献
93.
94.
By means of indirect double immunofluorescent staining,the coordination of PI antigen and perichromonucleolin(PCN),the constituent of nuclear periphery and nucleolus respectively,in the assembly and disassembly of chromosome pellicle during mitosis was studied.It was found that in 3T3 cells,during mitosis PI antigen began to coat the condensing chromosome surface earlier than PCN did.However,both of them completed their coating on chromosome at approximately the same stage of mitosis,prometaphase metaphase,The dissociation of mitosis,Prometaphase metaphase.The dissociation of PI antigen from chromosome pellicle to participate the formation of nuclear periphery took place also ahead of that of PCN,At early telophase PI antigen had been extensively involved in the formation of nuclear periphery,while PCN remained in association with the surface of decondensing chromosomes.At late telophase,when PI antigen was localized in an fairly well formed nuclear periphery,PCN was in a stage of forming prenucleolar bodies. 相似文献
95.
96.
J J Barycki L K O'Brien A W Strauss L J Banaszak 《The Journal of biological chemistry》2001,276(39):36718-36726
l-3-Hydroxyacyl-CoA dehydrogenase (HAD), the penultimate enzyme in the beta-oxidation spiral, reversibly catalyzes the conversion of l-3-hydroxyacyl-CoA to the corresponding 3-ketoacyl-CoA. Similar to other dehydrogenases, HAD contains a general acid/base, His(158), which is within hydrogen bond distance of a carboxylate, Glu(170). To investigate its function in this catalytic dyad, Glu(170) was replaced with glutamine (E170Q), and the mutant enzyme was characterized. Whereas substrate and cofactor binding were unaffected by the mutation, E170Q exhibited diminished catalytic activity. Protonation of the catalytic histidine did not restore wild-type activity, indicating that modulation of the pK(a) of His(158) is not the sole function of Glu(170). The pH profile of charge transfer complex formation, an independent indicator of active site integrity, was unaltered by the amino acid substitution, but the intensity of the charge transfer band was diminished. This observation, coupled with significantly reduced enzymatic stability of the E170Q mutant, implicates Glu(170) in maintenance of active site architecture. Examination of the crystal structure of E170Q in complex with NAD(+) and acetoacetyl-CoA (R = 21.9%, R(free) = 27.6%, 2.2 A) reveals that Gln(170) no longer hydrogen bonds to the side chain of His(158). Instead, the imidazole ring is nearly perpendicular to its placement in the comparable native complex and no longer positioned for efficient catalysis. 相似文献
97.
The principal absorptive cell of the rat small intestinal epithelium contains two homologous cytosolic proteins that bind long chain fatty acids. These are known as intestinal and liver fatty acid binding proteins (FABP). While their precise physiological roles have not been defined, they are believed to represent a multifunctional cytosolic transport system that is involved in the trafficking of exogenous lipids to sites of metabolic processing. 13C NMR studies have revealed differences in their fatty acid binding stoichiometries, binding mechanisms, and the ionization properties of bound fatty acids. To understand the functional differences, liver FABP has been crystallized for eventual comparison with the known crystal structure of intestinal FABP. The lattice type is trigonal with unit cell dimensions of a = b = 84.1 A and c = 44.2 A. The space group as determined by examination of the Patterson symmetry is either P3(1)21 or P3(2)21. 相似文献
98.
Z H Xu M K Buelt L J Banaszak D A Bernlohr 《The Journal of biological chemistry》1991,266(22):14367-14370
The murine adipocyte lipid binding protein (ALBP/aP2) has been cloned and expressed in Escherichia coli, purified to homogeneity, biochemically characterized, and crystallized for x-ray diffraction study. In the cloning, the ALBP coding region was placed under control of the recA promoter and downstream of the phage T7 g-10 translation enhancer sequence. Nalidixic acid (50 micrograms/ml) induced the expression of ALBP 20-fold over that attained using the pT7 system previously reported (Chinander, L. L., and Bernlohr, D. A. (1989) J. Biol. Chem. 264, 19564-19572). Recombinant ALBP was purified to homogeneity using a combination of pH fractionation, gel filtration, and immobilized metal affinity chromatography. The fluorescent affinity ligand 12-(9-anthroyloxy)oleic acid bound to homogeneous ALBP with an apparent Kd of 0.5 microM. rALBP was devoid of endogenous fatty acid, and oleic acid inhibited cysteine 117 modification by 5,5' -dithiobis-(2-nitrobenzoic acid) indicating integrity of the binding domain. Recombinant ALBP was phosphorylated by the soluble kinase domain of the insulin receptor with a Vmax of 11 nmol.min.mg of kinase and an apparent Km of 270 microM. Purified protein was crystallized using the hanging drop method with seeding. Crystalline ALBP was orthorhombic with cell dimensions of a = 34.4 A, b = 54.8 A, and c = 76.3 A. The space group was P212121, and there was one molecule per asymmetric unit. 相似文献
99.
100.
Kristina Hellberg Paul A Grimsrud Andrew C Kruse Leonard J Banaszak Douglas H Ohlendorf David A Bernlohr 《Protein science : a publication of the Protein Society》2010,19(8):1480-1489
Fatty acid binding proteins (FABP) have been characterized as facilitating the intracellular solubilization and transport of long‐chain fatty acyl carboxylates via noncovalent interactions. More recent work has shown that the adipocyte FABP is also covalently modified in vivo on Cys117 with 4‐hydroxy‐2‐nonenal (4‐HNE), a bioactive aldehyde linked to oxidative stress and inflammation. To evaluate 4‐HNE binding and modification, the crystal structures of adipocyte FABP covalently and noncovalently bound to 4‐HNE have been solved to 1.9 Å and 2.3 Å resolution, respectively. While the 4‐HNE in the noncovalently modified protein is coordinated similarly to a carboxylate of a fatty acid, the covalent form show a novel coordination through a water molecule at the polar end of the lipid. Other defining features between the two structures with 4‐HNE and previously solved structures of the protein include a peptide flip between residues Ala36 and Lys37 and the rotation of the side chain of Phe57 into its closed conformation. Representing the first structure of an endogenous target protein covalently modified by 4‐HNE, these results define a new class of in vivo ligands for FABPs and extend their physiological substrates to include bioactive aldehydes. 相似文献