Partial restoration of inactivated ribosomes with sodium borohydride or amino acids.
The aldehyde radical of ribose C1' at position 4324 in rat liver 28S rRNA generated by RNA N-glycosidase was either reduced to an hydroxyl group by sodium borohydride or converted into aldimine through a nucleophilic addition of amino acid used as a primary amine. Analysis of the R-fragment of 28S rRNA by polyacrylamide gel electrophoresis showed that the reduction of aldehyde to an hydroxyl group with sodium borohydride was highly specific. The protein synthesis activity of modified ribosomes was partially restored with the removal of the active aldehyde by sodium borohydride or amino acid. Reduction of aldehyde with sodium borohydride restored 43.1% of the protein synthesis activity. Among the twenty natural amino acids tested, tryptophan and histidine could restore 57.4% and 42.1% of the ribosome activity when brome mosaic virus RNA was used as mRNA. We came to the conclusion that the active aldehyde radical at position 4324 of 28S rRNA in modified ribosome may cause the inactivation of the ribosome for protein synthesis.
Li
Q
Q
Shanghai Institute of Biochemistry, Academiia Sinica, People's Republic of China.
Ling
J
J
Liu
W Y
WY
D020418:Algal Proteins; D000596:Amino Acids / Q000494:pharmacology*; D000818:Animals; D001894:Borohydrides / Q000494:pharmacology*; D002458:Cell Fractionation; D005609:Free Radicals / Q000378:metabolism; D005656:Fungal Proteins / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D011506:Proteins / Q000494:pharmacology; D012339:RNA, Ribosomal, 28S / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D054788:Ribosome Inactivating Proteins; D054790:Ribosome Inactivating Proteins, Type 2; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure
D020418:Algal Proteins; D000596:Amino Acids; D001894:Borohydrides; D005609:Free Radicals; D005656:Fungal Proteins; D011506:Proteins; D012339:RNA, Ribosomal, 28S; D054790:Ribosome Inactivating Proteins, Type 2; C061930:cinnamomin, Phytophthora cinnamomi; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins