Aminoacyltransferase II from Rat Liver

Peptidylpuromycin formation on mammalian polysomes. Transpeptidation and translocation

“…Kinetic Studies of Transpeptidation. In our previous reports (Schneider et al, 1968;Raeburn et al, 1971), the reaction of nascent polypeptide chains with puromycin was utilized primarily for studying translocation catalyzed by EF-2. Advantages of the system apply equally to the study of peptidebond formation per se.…”

“…[3H]NAD+ and diphtheria toxin under conditions which assure complete reaction of EF-2 (Raeburn et a!., 1971). The transfer of [14C]leucine from [14C]leucyl-tRNA into protein was carried out as described (Raeburn et al, 1971).…”

Isolation and distribution of elongation factor 2 in eggs and embryos of sea urchins

“…This protein factor can be studied by taking advantage of its characteristic property of ADPribosylation in the presence of diphtheria toxin and NAD. The toxin catalyzes the transfer of one moiety of ADP-ribose from NAD to one molecule of EF-2, providing an easy and accurate means of determining the amount of EF-2 (Goor & Maxwell, 1970;Raeburn et al, 1971;Pappenheimer, 1977;Bermek, 1978). In this paper, we determined the quantity and subcellular localization of EF-2 in eggs and embryos.…”

“…Quantitation of Elongation Factor 2. The content of in fractions obtained by differential centrifugation and by sucrose gradient fractionation was assayed by using the diphtheria toxin dependent ADP-ribosylation of EF-2 (Goor & Maxwell, 1970;Raeburn et al, 1971;Yablonka-Reuveni & Warner, 1979;Yablonka-Reuveni et al, 1983).…”

“…The Filters were oven dried at 80 °C for 10 min and dissolved in 10 mL of Beckman Ready Solv HP scintillant, and their radioactivity was measured in a Beckman LS 8000 scintillation counter. The concentration of EF-2 was determined from the extent of [3H]-ADP-ribosylation by assuming that one molecule of ADPribose is bound to one molecule of EF-2 in the presence of diphtheria toxin and excess NAD (Raeburn et al, 1971).…”