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Biotin uptake has been investigated using an Escherichia coli biotin requiring auxotroph grown under biotin-deficient conditions. This strain accumulated biotin in the free and bound form. In agreement with a previous report by O. Prakash and M.A. Eisenberg (J. Bacteriol. 120 (1974) 785–791), the biotin entry proved to be an active process which depended on an energy source and was inhibited in the presence of uncouplers. The kinetic parameters have been determined (KM = 0.05 μM, Vmax = 7 pmol/min per mg dry weight). The pool of free biotin could be readily exchanged with external biotin and decreased to a very low level in the absence of an energy source. The use of several biotin analogues revealed that this transport system was quite specific for biotin: slight modifications, for instance in the valeric chain. lowered drastically the affinity for the carrier. 相似文献
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The influence of the trichloroacetyl group on the 13C chemical shift of the substituted and vicinal carbon atoms allows the unambiguous assignment of the signals of the carbon atoms. For derivatives of 1,2,3,4,6-penta-O-acetyl-β-d-glucopyranose and β-gentiobiose octaacetate, the “α-effect” is +4 to +5 p.p.m. (deshielding), whereas the “β-effect” has smaller and always negative values. 相似文献
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Preferential reassociation of immunoglobulin H and L chains was investigated using a method of competitive hybridization. A model was built in which two monoclonal light chains, one autologous, LA, and one heterologous, LB, were competitively reassociated with a given monoclonal heavy chain, HA. A total of 12 human myeloma proteins were used with known isotypic, allotypic and variability subgroups: 44 distinct combinations were studied by competitive hybridizations. It was found that a preferential reassociation occurred between the complementary H and L chains that were associated in the native molecule in 80% of the cases. It was clearly established that the subgroups had no influence on the preferential reassociations that seem, therefore, to rely exclusively on individual (“idiotypic”) structural differences. It was shown that, although H and L chains had been fully reduced and denatured, the same degree of preferential reassociation was observed after the chains had been reoxidized and refolded. These experiments suggest, therefore, that the observed preferential reassociations are the consequence of an antigen-independent selection process that must have taken place during the differentiation of the antibody-forming cells. 相似文献
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Eric G. Brunngraber Javaid I. Javaid 《Biochimica et Biophysica Acta (BBA)/General Subjects》1975,404(1):67-73
Mannose-rich glycopeptides derived from brain glycoproteins were obtained by proteolysis of bovine brain tissue or subcellular fractions derived from rat brain tissue. The dialyzable mannose-rich glycopeptides were isolated by colum electrophoresis and gel flitration. These glycopeptides contained, on the average, six mannose and two N-acetylglucosamine residues with variable amounts of fucose and galactose. Over 50% of the mannose-rich glycopeptides of rat brain were localized in the microsomal and synaptosomal fractions; myelin and the soluble fraction contained lesser amounts. None was recovered from the mitochondria. The amount, per mg protein, of mannose-rich oligosaccharide chains in the myelin exceeded the concentration found in the microsomal and synaptosomal fractions. The concentration of mannose-rich glycopeptides derived from glycoproteins was 50% higher in white matter than in gray. On the other hand, the non-dialyzable and acidic sialoglycopeptides showed a three-fold enrichment in gray matter compared to white. The relatively lower ratio of sialoglycopeptides to mannose-rich glycopeptides observed in white matter (2.5) compared to gray matter (6.9) is reflected in the lower value for the ratio in myelin (1.1) compared to synpatosomes (2.1). Although glycoproteins that contain mannose-rich oligosaccharide chains are present in the nerve cell and its terminals, these glycoproteins appear to be relatively enriched in myelin and/or glial membranes. 相似文献