Crystallographic studies on the structure and function of glycogen phosphorylase b.
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Biomedical subjects
Publications and source records attributed to L N Johnson.
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The conformations of lysozyme in crystals and in aqueous solution are discussed and it is shown that the basic conformation is similar in the two states. Certain parts of the molecule have mobility. The reactions of lysozyme with protons, metal ions and some organic reagents are examined in the light of the conformations and their dynamics. The reactions considered are mainly those of tyrosyl, tryptophyl and carboxylate residues. The reactivity data are used in a discussion of the energy states of the reacting side-chains. In particular the reactivity of Glu-35 and its interaction with Trp-108 lead to suggestions for some new aspects in the hypothesis for the mechanism of action of lysozyme. In most respects the X-ray crystal diffraction and the nuclear magnetic resonance solution studies are in accord and complementary.
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1. The interaction of a variety of fluorescent probes with the membranes of adrenal medullary chromaffin granules is described. 2. Changes in the motional properties of the bound probes with temperature were investigated and evidence is presented which indicates that ordering of the membrane lipids occurs below 33 degrees C. 3. The ordering is characteristics of the membrane lipids and is retained by sonicated aqueous dispersions of the total lipid extracted from chromaffin granule membranes. 4. The ATPase and NADH:acceptor oxidoreductase activities of the chromaffin granule membrane have discontinuous Arrhenius temperature versus activity relationships with 'transitions' at 33 degrees C. 5. The ATPase has a second transition at 36.5 degrees C. 6. The 33 degrees C 'transition' for the NADH:acceptor oxidoreductase is removed by treatment with the detergent Triton X-100. 7. The correlation between the onset of lipid ordering and the change in activation energy of the membrane-bound enzyme activities is discussed in terms of the co-operative interactions of the different membrane components. The possible role of lipid ordering in exocytosis is discussed.
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The distribution of concanavalin A (con A) receptor sites on the membranes of chromaffin granules has been investigated by binding studies using 125I-labelled con A and by electron-microscope studies using ferritin-labelled con A. In both experiments con A was observed to bind to chromaffin granule membranes but not to intact granules. The ferritin-con A particles bind to only one of the two possible surfaces of the chromaffin granule membranes. These results are in agreement with previous observations concerning the asymmetric distribution of saccharide residues on the surfaces of a number of different plasma membranes. They suggest that for the intracellular membrane of the chromaffin granule the saccharide sites, like those in plasma membranes, are not exposed to the cell cytoplasm. Further work is necessary to establish whether these sites are on the inner surface of the membrane or whether they are unmasked during the conversion of granules to membrane ghosts.
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