Procedure for determination of participation of aminoacyl-tRNA in label distribution in studies on protein degradation in vivo.
The determination of radioactive amino acid incorporation into proteins is important in measurements of protein degradation. Aminoacyl-tRNAs participating in label distribution may interfere during these measurements. To check this possibility, a separate procedure with ion-exchange paper DE 81 allowing the percentage of radioactivity in aminoacyl-tRNA to be determined was developed. After application of microliter amounts of Trametes versicolor mycelium extracts on DE 81 paper circles, stepwise elution with buffers of different pH's and ionic strengths was carried out. This enabled labelled proteins to be separated from labelled aminoacyl-tRNAs and unbound amino acids.
D002852:Chromatography, Ion Exchange; D002854:Chromatography, Paper; D005656:Fungal Proteins / Q000032:analysis*; D011124:Polyporaceae; D012346:RNA, Transfer, Amino Acyl / Q000032:analysis*
D005656:Fungal Proteins; D012346:RNA, Transfer, Amino Acyl