Abstract
This chapter describes the purification of IF-I and of IF-II from 0.5 M KC1 extracts of rabbit reticulocyte polysomes and several assays for the biological activities of each initiation factor. One function of IF-I during initiation complex formation is the binding of [ 35 S]Met-tRNA i Met to a 40 S ribosomal subunit in a GTP-dependent but AUG-, AUG(U) 25 -, and mRNA-independent reaction. IF-I also forms a ternary complex with [ 35 S] Met-tRNA i Met and GTP; this complex is retained by Millipore filters. This retention property forms the basis of a convenient and routine assay for the presence of IF-I activity.
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