Affinity electrophoretic detection of primary amino groups in nucleic acids: application to modified bases of tRNA and to aminoacylation.
Thiolation of primary amino groups in tRNA with the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate gives rise to species which are retarded during electrophoresis in organomercury-containing polyacrylamide gels. Since such amino groups occur, as far as is known, only as part of the modified bases 3-(3-amino-3-carboxypropyl)uridine and N-2-(5-amino-5-carboxypentyl)cytidine or as the alpha-amino group of aminoacylated tRNAs, this extension of the principle of affinity electrophoresis can be used for the detection and analysis of a specific functional group in both single tRNA species and in a mixed population. The strength of the interaction may be quantified and provides information on the chemical environment/conformation of the derivatized bases.
Igloi
G L
GL
Institut für Biologie III, Universität Freiburg, Germany.
D000588:Amines / Q000032:analysis*; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry; D012343:RNA, Transfer / Q000737:chemistry* / Q000302:isolation & purification; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry* / Q000302:isolation & purification; D012352:RNA, Transfer, Glu / Q000737:chemistry; D012357:RNA, Transfer, Lys / Q000737:chemistry; D012358:RNA, Transfer, Met / Q000737:chemistry; D013388:Succinimides; D013439:Sulfhydryl Reagents; D003313:Zea mays / Q000737:chemistry
D000588:Amines; D012346:RNA, Transfer, Amino Acyl; D012352:RNA, Transfer, Glu; D012357:RNA, Transfer, Lys; D012358:RNA, Transfer, Met; D013388:Succinimides; D013439:Sulfhydryl Reagents; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D012343:RNA, Transfer