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BIOL300%20Lecture%204%20S.H.

BIOL300%20Lecture%204%20S.H. - Make a non-phosphorylatable...

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Unformatted text preview: Make a non-phosphorylatable inactive protein: How to analyze the function of kinase substrates Make an always “phosphorylated” protein by changing to phosphomimetic amino acid-These approaches are necessary in vitro as well as in vivo.-In vitro: e.g. incubation with radio-labelled ATP and peptide analysis with proteases of proteins with known sequences.-In vitro: mass spectrometry-In vivo to determine if events really happen in vivo, and their significance: functional testing. (E) Autocrine signaling allows cells to behave as a group Used by cells in tissue culture; Also used by cancer cells to overcome external controls High K d means fast off rate, that is quick termination possible, but high [L] required, e.g. synaptic signaling Dominant/recessive Gene/protein cascades Mutant analysis Genetic analysis 16_08_cascades.jpg ? ? ? Wild-type shy m/m shy +/m Blush +/M Blush M/M Blush +/M shy +/m Double mutant Lecture 4 G protein-coupled receptor signaling I Reading for Lectures 4 - 5: Lodish Ch. 15, pp. 635-660 G protein-coupled receptors (GPCRs) are seven-pass transmembrane receptors....
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BIOL300%20Lecture%204%20S.H. - Make a non-phosphorylatable...

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