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- W933842085 abstract "Publisher Summary This chapter discusses the measurement of stoichiometry of protein phosphorylation by biosynthetic labeling. Determination of the extent of phosphorylation of proteins that can be obtained only in trace amounts or in impure form is not possible by chemical means. The stoichiometry of phosphorylation of such proteins can be inferred if the protein of interest can be labeled biosynthetically. This is accomplished by: (1) labeling the cells to equilibrium with a radioactive amino acid and with [ 32 P]P i , (2) measurement of the specific activity of total cellular protein and intracellular adenosine triphosphate (ATP), (3) isolation of the protein of interest by immunoprecipitation, (4) quantification of the protein from the recovery of amino acid label, and (5) calculation of the number of moles of phosphate present in the protein from the recovery of 32 P. The specific activity of the cellular ATP pool has been estimated to reach equilibrium with that of the medium in as little as 6 hours after the addition of medium containing [ 32 P]P i to cells." @default.
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- W933842085 date "1991-01-01" @default.
- W933842085 modified "2023-10-11" @default.
- W933842085 title "[20] Measurement of stoichiometry of protein phosphorylation by biosynthetic labeling" @default.
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- W933842085 doi "https://doi.org/10.1016/0076-6879(91)01022-t" @default.
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