Biomedical subjects
Publications and source records attributed to S R Ayad.
In this paper we show that the measurement of cyclic AMP by a protein saturation assay deviates from ideality due to the presence of two classes of binding sites. An equation was derived which completely describes saturation assays and the effects on these of varying each of the component parameters was measured. The relevance of these findings to other protein saturation assays is discussed.
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Hybrids (PCM) between a malignant mouse lymphoma suspension cell line (P388F-36) and a normal Chinese hamster fibroblastic cell line (Ch23) have already been isolated in this laboratory. Investigations were carried out on the cAMP phospodiesterases of the parents and two of these hybrids--PCM2 and PCM3. PCM3 shows a rather unusual growth characteristic in that a considerable proportion of the cells exist at any one time either in suspension or only loosely attached to the substratum, the remaining cell population existing in a monolayer form. It was found that each cell line exhibited multiple forms of the enzyme with varying affinities for cAMP. Both parents, although different, contained high-, low-, and extra-high apparent Km forms of the enzyme. The hybrids exhibited characteristics of both parental systems but were different from each other. Neither hybrid exhibited a high-Km enzyme, but both exhibited two low-Km forms. There was also a slight variation between monolayer and suspension cells of PCM3 hybrid. An attempt has been made to explain these phenomena with respect to hybridization and the growth characteristics of the cells.
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1. The electrophoretic mobilities of a mouse lymphoma cell, a Chinese-hamster fibroblast and a somatic-cell hybrid (also fibroblastic), produced by fusion of the hamster cell and a mouse lymphoma cell, were measured at 25 degrees C over a range of pH, concentration of Ca2+ ions and concentration of La3+ ions. 2. All the cells have pI at pH3.5. 3. Ca2+ ions decrease the mobilities and zeta potentials of the cells to zero in the range 1-100mM. 4. La3+ ions lower the mobilities and zeta potentials in the range 10 muM-1 mM, and the cells become positively charged above 1 mM. 5. The data are consistent with specific adsorption of La3+ ions on approx. 2 X 10(14) sites/m2 of cell surface with a free energy of approx. -37kJ/mol. 6. The effects of Ca2+, La3+ and ionic strength on the extent of aggregation of the cells and of neuraminidase-treated cells were studied. 7. Ca2+ ions do not markedly increase aggregation, whereas La3+ ions gave rise to extensive aggregation in the range 10 muM-1 mM, corresponding to the region of La3+ adsorption. 8. Both fibroblastic cell lines are aggregated at high ionic strength. 9. The fibroblastic cells have larger amounts of trypsin-sensitive carbohydrate than does the lymphoma cell; the possible role of this material in cellular aggregation is discussed.