G. Hortin, I. Boime
Methods in enzymology
Publisher Summary This chapter presents general considerations of applications of amino acid analogs for studying co- and post-translational modifications of proteins. This approach involves selection of amino acid analogs, selection of analog concentration, demonstration of amino acid analog incorporation into protein, and analysis of the effects of amino acid analogs on protein processing. When a particular analog is applied to a new system, it is necessary to assess critically whether the analog is incorporated into protein and to establish its effective concentration. Selection of amino acids for incorporation into protein depends on the selectivity of aminoacyl-tRNA synthetases, which possess an extraordinary ability to discriminate among structurally similar amino acid substrates. However, determination of the appropriate analog concentration for use with intact cells presents greater difficulty owing to the added complications of uptake into cells, amino acid metabolism, and endogenous amino acid pools.