Influence of cholesteryl 14-methylhexadecanoate on some ribosomal functions required for peptide elongation.
1. Polyribosomes and ribosomal subunits from rat liver were adsorbed on a cellulosic ion-exchange adsorbent, freeze-dried and extracted with organic solvents. The activity of extracted particles in peptide elongation was tested in the presence of purified peptideelongation factors. 2. Chloroform-methanol mixture (2:1, v/v) extracted 1.87+/-0.15 pmol of cholesteryl 14-methylhexadecanoate/pmol of the smaller ribosomal subunit and 0.92+/-0.11 pmol/pmol of the larger subunit. 3. In the presence of transferase I, extracted polyribosomes and 40S subunits bound more phenylalanyl-tRNA than did control non-extracted particles. The same binding as in control mixtures was obtained with extracted particles supplemented with cholesteryl 14-methylhexadecanoate in quantities corresponding to those extracted. 4. The polymerization of phenylalanine was greatly decreased with extracted polyribosomes and subunits and addition of the cholesteryl ester could not fully restore the original activity. 5. Extraction significantly decreased the activity of the P site of peptidyl transferase and normal activity was recovered after the addition of the ester. The A site of peptidyl transferase in extracted polyribosomes showed an increased activity when compared with non-extracted polyribosomes. 6. Cholesteryl 14-methylhexadecanoate apparently affects the function of the ribosomal A site and peptidyl transferase site and probably also that of the guanosine triphosphatase site and P site. The presence of different amounts of the ester in polyribosomes may be one of the mechanisms modulating peptide elongation at the ribosomal level.
D000818:Animals; D002501:Centrifugation, Zonal; D002784:Cholesterol / Q000494:pharmacology*; D002788:Cholesterol Esters; D002852:Chromatography, Ion Exchange; D004926:Escherichia coli; D005227:Fatty Acids / Q000494:pharmacology*; D005260:Female; D006150:Guanine Nucleotides; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D010453:Peptide Synthases / Q000378:metabolism; D010649:Phenylalanine; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism; D014176:Protein Biosynthesis; D012329:RNA, Bacterial; D012343:RNA, Transfer; D051381:Rats; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*; D014316:Tritium
D002788:Cholesterol Esters; D005227:Fatty Acids; D006150:Guanine Nucleotides; D012329:RNA, Bacterial; D014316:Tritium; C005463:carcinolipin; D010649:Phenylalanine; D012343:RNA, Transfer; D002784:Cholesterol; D010453:Peptide Synthases