The use of synthetic tRNAs as probes for examining nascent peptides on Escherichia coli ribosomes.
The polyuridylic acid-dependent syntheses of polycysteine and polyserine were carried out on Escherichia coli ribosomes using two new synthetic tRNA species. The peptides were initiated with N-acetyl or N-acyl coumarin derivatives of either Ser-tRNA or Phe-tRNA. The properties of the resulting nascent peptides were compared to those of nascent polyphenylalanine chains synthesized under similar conditions. This was accomplished by following changes in the fluorescence properties of the probes covalently linked to the amino-terminus of each of the nascent polypeptides as they were formed on the ribosomes. Nascent polycysteine and polyserine peptides appeared quite different from those of polyphenylalanine, as indicated by the anisotropy of fluorescence from the amino terminal probe. In contrast to serine and cysteine peptides, the synthesis of all the polyphenylalanine peptides was insensitive to inhibition by erythromycin, even though these peptides were initiated with N-acyl serine. The results support the hypothesis that nascent polyphenylalanine peptides have atypical physical and chemical properties and demonstrate the utility of using modified tRNAs to study ribosome function and the synthesis of proteins.
Picking
W
W
Department of Chemistry and Biochemistry, University of Texas, Austin 78712.
Picking
W D
WD
Hardesty
B
B
D001483:Base Sequence; D004926:Escherichia coli / Q000378:metabolism; D008969:Molecular Sequence Data; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry; D011072:Poly U / Q000378:metabolism; D011487:Protein Conformation; D015347:RNA Probes; D012343:RNA, Transfer / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D010455:Peptides; D015347:RNA Probes; C027794:polyserine; D011072:Poly U; C012001:polyphenylalanine; C057713:polycysteine; D012343:RNA, Transfer