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71.
Controlled acid hydrolysis of poly-(2,8)-linked homopolymers ofN-acetylneuraminic acid (colominic acid) and its homologous poly-N-glycolylneuraminic acid afforded high yields of the corresponding disaccharides useful in block synthesis of disialylated gangliosides. The poly-N-glycolylanalog was derived from de-N-acetylated colominic acid by two different reaction sequences. The first one involved reaction with acetoxyacetyl chloride followed by de-O-acetylation. The second and most interesting one requiredN-acryloylation and reductive ozonolysis. 相似文献
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74.
High resolution 1H-n.m.r. study of the interaction between initiation factor IF1 and 30S ribosomal subunits
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Interaction between Escherichia coli translational initiation factor IF1 (mol. wt. 8119) and 30S ribosomal subunits was followed by high resolution 1H-n.m.r. spectroscopy. Upon gradual addition of increasing yet largely sub-stoichiometric amounts of biologically active deuterated 30S ribosomal subunits, selective line broadenings and chemical shift changes were observed against the background of the gradual disappearance of the whole spectrum. At the highest 30S:IF1 ratio attained (0.25), all the resonance lines were broadened beyond meaningful detection. This behaviour, which can be partly reversed by increasing the ionic strength and/or the temperature, is due to the interaction between IF1 and the 30S ribosomal subunits, and can be explained by the existence of a medium-fast exchange dynamics between free and bound factor. The selective effects observed during titration with 30S ribosomal subunits shed some light on the mode of interaction of IF1 with 30S ribosomal subunits. At least one of the two His residues of the factor appears to be involved in the binding, since it undergoes a low-field change of chemical shift and becomes totally immobilized in the IF1-30S complex. Also strongly implicated in the interaction with 30S are more than one Ser and Arg residue and probably one lysine. Additional effects of the interaction of IF1 with ribosomes are a drastic reduction in the intensity of the ring current upfield shifted methyl resonances and mobilization of a previously rotationally hindered phenylalanine ring. 相似文献
75.
The chemical synthesis of oligoribonucleotides VII. A comparison of condensing agents in the coupling of silylated ribonucleosides. 总被引:2,自引:2,他引:0
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The t-butyldimethylsilyl group is shown to be an ideal protecting group for the 2T-hydroxyl function of ribonucleosides during the synthesis of ribonucleotides using any of nine commonly used condensing agents. The phosphite coupling procedure compares favorably with all of the widely used condensing agents and provides a most convenient route to the key intermediates in the "modified" triester strategy. 相似文献
76.
The interaction of thioformyldistamycin, an amide isostere of the naturally occurring antibiotic distamycin A, with a self-complementary decadeoxynucleotide duplex, d(CGCAATTGCG)2, has been examined using a variety of high-field 1H-NMR techniques. The ligand exhibits two forms in solution arising from geometric isomerism due to restricted rotation around the thioformamide bond. Only the thermodynamically more stable Z-form is shown to bind to the oligonucleotide along its minor groove at the central 5'-AATT segment with the end groups of the ligand extending into the flanking GC regions but without any close contact at the amidinium terminus. Cross-peaks involving characteristic intra- and interresidue proton connectivities in the 2D experiments (COSY and NOESY) were employed to assign individual resonances of both strands in the asymmetric DNA-drug complex. The solution structure of the complex was constructed by molecular mechanics calculations based upon initial estimates of drug-DNA NOE contacts and further refined through energy minimization. These results complement previous structural studies on distamycin and other lexitropsins with oligonucleotides. The exchange of the ligand between two equivalent binding sites on the DNA sequence was estimated to occur at 40 s-1 with a free energy of activation of 16.5 kcal.mol-1 at 321-326 K. There was no evidence of formation of a 2:1 drug-oligomer complex, in contrast to the case of the natural product, which is attributed to steric demands of the larger sulfur atom. 相似文献
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78.
Translational initiation factor-3 (IF-3) was characterized by 1H NMR spectroscopy as a function of pH and temperature and following chemical modifications. Spin-lattice relaxation times for individual resonances and bands were also measured. Several resonances were assigned to different amino acid residues by different criteria. Among these are the CH3-N of the N-terminal methionine which appears free, mobile, and very sensitive to the modification of several physicochemical parameters as well as the 3,5 and 2,6 protons of the three tyrosines (two of which play a role in the function of IF-3) which were found to be located in different magnetic environments. Two of these residues appear to be close to each other and in the vicinity of a slow reacting arginine within the tertiary structure of the factor. The properties and the titration behavior of the imidazole proton resonances suggest that the single His residue is partially buried in the protein structure. Characteristic of the IF-3 spectrum also is the presence of an abundant subset of Arg delta-CH2, Lys epsilon-CH2, and CH3 protons displaying clear cut upfield perturbations. These are probably due to the coming together of two or more apolar "fronts" which possibly arise from distant parts of the molecule and result in the close proximity between aromatic rings and aliphatic side chains. The IF-3 spectrum also includes several distinct methyl resonances significantly shifted upfield by aromatic ring currents. Overall, the characteristics of the spectrum, its relative insensitivity to temperature and ionic strength, and the existence of extensive cross-relaxation phenomena indicate that IF-3 has a highly folded tertiary structure with abundant hydrophobic regions. In spite of some heterogeneity in the distribution of the side chain environments, no indication was found for the existence of distinct domains or, at least, of extensive regions with higher mobility. 相似文献
79.
Proteins from the prokaryotic nucleoid. High-resolution 1H NMR spectroscopic study of Escherichia coli DNA-binding proteins NS1 and NS2 总被引:1,自引:0,他引:1
The 1H-NMR spectra of the two Escherichia coli basic, low-Mr (approximately equal to 9000) DNA-binding proteins NS1 and NS2 and of their native complex NS were studied at 400 MHz and a number of resonances and resonance peaks were assigned. As in the case of some eukaryotic histones, the presence of a large number of high-field perturbed Phe resonances, several shielded and deshielded methyl resonances and backbone NH protons quite inaccessible to the solvent clearly indicate the existence of extensive tertiary and, even more so, quaternary structures involving hydrophobic interactions. These structures are lost upon heating, but readily reform upon cooling. Spectral differences between NS1, NS2 and NS and the greater thermal stability of NS indicate that molecules of the heterologous subunits (NS1 and NS2) aggregate (dimerize) preferentially in comparison to the self-aggregation of the homologous subunits. Unlike those of the eukaryotic histones, the tertiary and quaternary structures of NS are insensitive to extensive variations of the ionic strength. 相似文献
80.
This study examined the changes in protein phosphorylation in response to cholinergic (muscarinic) stimulation of salivary secretion in the rat submandibular gland. Carbachol stimulation was associated with phosphorylation in a number of protein bands as detected by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and autoradiography. The molecular masses (Mr) of two proteins, in which the amount of phosphorylation more than doubled in response to carbachol, were 22,000 and 96,000. The Mr 96,000 protein precipitated at 120,000 X g while most of the Mr 22,000 protein remained in the supernatant at this speed. The effect of carbachol on the phosphorylation of the Mr 22,000 and 96,000 proteins was blocked by atropine, indicating that the cholinergic receptor involved is muscarinic. The time course of phosphorylation of the Mr 22,000 protein consisted of a rapid increase in phosphorylation within the first min of carbachol stimulation. This increased phosphorylation persisted for less than 1 min. The increased phosphorylation of the Mr 96,000 protein also occurred within the first min but it persisted for at least 10 min. However, removal of the muscarinic agonist, carbachol, resulted in the rapid dephosphorylation of this protein. When the plasma membranes were purified, the Mr 96,000 protein was phosphorylated by ATP in the presence of Na+ and Mg2+. It was dephosphorylated by K+. This proves that the Mr 96,000 dalton protein is the alpha-subunit of the (Na+ + K+)-ATPase. 相似文献