Synthesis of thiol-containing analogues of puromycin and a study of their interaction with N-acetylphenylalanyl-transfer ribonucleic acid on ribosomes to form thioesters.
1. The thiol-containing analogue of puromycin, 6-dimethylamino-9-{1'-[3'-(2''-mercapto-3''-phenylpropionamido)-3'-deoxy-beta-d-ribofuranosyl]}purine (XVII) in which the primary amino group of the antibiotic is replaced with a thiol grouping, was synthesized chemically (compound XVII is abbreviated to thiopuromycin). 2. Thiopuromycin (XVII) was found to be active in releasing N-[(3)H]acetylphenylalanine from its tRNA carrier as the thioester, N-acetylphenylalanylthiopuromycin (XIX) in the Escherichia coli ribosomal system. The reaction product (XIX) was synthesized chemically from thiopuromycin and N-acetylphenylalanine and found to be stable to hydrolysis in the standard incubation medium at pH7.6. dl-Phenyl-lactylpuromycin (XXI), the hydroxy analogue of puromycin, was also synthesized chemically and shown to release N-acetylphenylalanine from its tRNA carrier in the E. coli ribosomal system, thus confirming the previous results of Fahnestock et al. [Biochemistry (1970) 9, 2477-2483]. 3. In marked contrast with the results obtained in the E. coli system, both thiopuromycin (XVII) and hydroxypuromycin (XXI) were found to be inactive in releasing N-acetylphenylalanine from its tRNA carrier in the rat liver ribosomal system.
D000818:Animals; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives; D014176:Protein Biosynthesis; D011691:Puromycin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D051381:Rats; D012270:Ribosomes / Q000378:metabolism*; D013045:Species Specificity; D013438:Sulfhydryl Compounds / Q000138:chemical synthesis / Q000378:metabolism
D012335:RNA, Ribosomal; D013438:Sulfhydryl Compounds; D010649:Phenylalanine; D011691:Puromycin; D012343:RNA, Transfer