PubMed · 927191
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G E Tarr. 1977. Improved manual sequencing methods.. https://doi.org/10.1016/0076-6879(77)47036-8
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D-Mannooligosaccharides (dp 1 to > 17) were released by mild acid hydrolysis from the phosphopeptidomannan of a Candida albicans strain of A serotype (VW.32). Among these, mannooligosaccharides ranging from bi- to hepta-ose, which were obtained in appreciable amounts, were structurally investigated and found to belong to the beta-D-(1-->2)-linked series. The occurrence of such compounds has already been reported in other Candida albicans strains. The complete 1H- and 13C-resonance assignments for manno-tri- to manno-hepta-ose are reported and general rules applicable for the 1NMR spectrum analysis of linear mannooligosaccharide of the general structure, beta-D-Man p-(1-->2)-[beta-D-Man p-(1-->2)]n-beta-D-Man p are proposed.
E. coli carbamyl phosphate synthetase binds 0.2-0.4 mol equivalents of glutamine in an acid resistant form. The bound material is quantitatively released as glutamate by weak base hydrolysis and as a mixture of 12% glutamate, 10% gamma-glutamylhydroxamate, and 70% pyrrollidonecarboxylic acid by hydrolysis with hydroxylamine. These results provide direct evidence for a gamma-glutamyl acyl ester on the enzyme. The absence of the acyl ester in a mutant carbamyl phosphate synthetase with a Cys269-->Ser substitution in the glutaminase subunit further suggests that the covalent intermediate is a thioester of Cys269. Under equilibrium conditions, the Cys269Ser mutant enzyme binds glutamine with a Kd of 7 +/- 1 microM, indicating that Cys269 is essential for acyl ester formation but not for binding of glutamine.
A method is described for the characterization of immune complex components by dot blot analysis. After isolation by chromatographic techniques and precipitation with polyethylene glycol, immune complexes were dissociated in 0.1 M phosphate (pH 2) and bound to a nitrocellulose membrane in a dot blot unit. Biotinylated probes were then used to identify the following immune complex components: specific antigens, biologically active antibodies, antibody isotypes, antibody subclasses, antibody idiotypes, and rheumatoid factors. This nonradioactive procedure takes less than 2 h to perform and has been used to analyze immune complexes isolated from sera (rabbit and human) and synovial fluid (human).