Biology - Molecular Biology In Medicinal Chemistry by Thomas Lengauer, Raimund Mannhold, Hugo Kubinyi, Hendrik

By Thomas Lengauer, Raimund Mannhold, Hugo Kubinyi, Hendrik Timmerman

Presents a vast, application-oriented assessment of this expertise. The state of the art in bioinformatics is evaluated both from a world view by way of introducing genuine program situations.

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The amount of fluorescence correlates directly to oxygen consumption in the well. This, in turn, relates to the number or relative viability of cells in this particular well. Any reaction that can be linked to oxygen consumption or generation can potentially be measured by using this system (Fig. 9). The signal is dynamic and is read in real-time. Samples may be read as frequently and as many times as desired. The signal is completely reversible, and will rise as the cells proliferate and then fall as they die, giving a more complete picture of proliferation and toxicological kinetics.

In addition microscopic applications allow analysis of intracellular spatial distributions which is not possible with flow cytometry. Mitotic cells are characterized by an increase in phosphorylated nucleolin, a nuclear protein phosphorylated in cells entering mitosis. Using a monoclonal antibody that specifically recognizes phosphorylated nucleolin, Mayer et al. [91] have developed a high-throughput whole-cell immunoblot assay. Compounds arresting cells in mitosis by novel modes of action could be identified after sorting compounds with secondary assays.

These experiments revealed that Cdc25 inhibition increased Erk phosphorylation and nuclear accumulation, and indicated that the phosphorylation status of Erk is regulated by a Cdc25 phosphatase. A phospho-histone H3 antibody that targets the phosphorylation site on Ser10 of histone H3 and a phospho-histone H1 antibody were used to detect mitogen- and oncogene-stimulated phosphorylation in small nucleosomal regions that are sensitive to hyperacetylation [88, 89]. Active kinase states have been analyzed with a panel of phosphospecific mono- 25 26 1 Cellular Assays in Drug Discovery clonal anti-kinase antibodies.

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