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

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

Offers a huge, application-oriented assessment of this know-how. The cutting-edge in bioinformatics is evaluated both from a world view by way of introducing actual program eventualities.

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This assay has been developed as a HTS assay technology by PerkinElmer Life Sciences. 6 Protein Fragment Complementation Assays (PCA) The methods described above to study protein–protein interactions are associated with certain shortcomings. The yeast two-hybrid system is based on transcription, which requires proteins to be available in the nucleus. In FRET, fluorescent proteins need to be expressed in cells at relatively high levels and the interaction must be such that the proteins are in close proximity to allow energy transfer.

5 GPCR Activation using Bioluminescence Resonance Energy Transfer (BRET) BRET is a naturally occurring process. The jelly fish A. victoria or the sea pansy Renilla reniformes create green fluorescence by BRET. In the former blue-lightemitting case, aequorin associates with GFP, whereas in the latter case, the energy generated by degradation of coelanterazine by luciferase is transferred to GFP. This transfer, however, only occurs if luciferase and GFP form a heterodimer. BRET has been used to demonstrate the interaction of circadian clock proteins from cyanobacteria expressed in E.

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.

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