Raman spectra and conformational properties of ribosomes during various stages of disassembly.
Raman spectra have been obtained on aqueous solutions of ribosomes, ribosomal subunits, ribosomal proteins, and ribosomal RNA extracted from both rat liver (RL) and Escherichia coli cells. The species examined from RL ribosomes are total ribosomes (80 S), large subunits (60 S), small subunits (40 S), EDTA-treated ribosomes, total rRNA, 28S RNA, 18S RNA, total protein, RNP particles from 80S ribosomes, and RNP particles from 60S subunits. The species examined from E. coli ribosomes are total ribosomes (70 S), large subunits (50 S), small subunits (30 S), mixtures of 50S and 30S subunits, total rRNA, 23S RNA, and 16S RNA. All rRNA molecules are shown by Raman spectroscopy to contain highly ordered secondary structures in which the backbone conformations are predominantly of the A-helix type. The present Raman spectra do not contain sufficient detail, however, to reach firm conclusions about the conformations of ribosomal proteins or their mutual interactions. RNA molecules within ribosomal particles remain highly ordered during various stages of ribosome disassembly, and their conformations are generally invariant to perturbations of ribosome structure, including dissociation into subunits, EDTA treatment, and partial deproteinization in a CsCl density gradient. However, when total protein extraction is carried out on ribosomes and subunits, small but significant changes in rRNA secondary structure are detected. The kind and magnitude of secondary structural change are different for different ribosomal particles. The Raman spectra of the ribosomes are compared also with spectra of a model ribonucleoprotein, the complex formed by poly(riboadenylic acid) and poly(L-arginine).
D000818:Animals; D002458:Cell Fractionation; D004926:Escherichia coli / Q000648:ultrastructure*; D008099:Liver / Q000648:ultrastructure*; D008954:Models, Biological; D009690:Nucleic Acid Conformation; D010455:Peptides; D011061:Poly A; D011487:Protein Conformation; D051381:Rats; D012261:Ribonucleoproteins / Q000032:analysis; D012270:Ribosomes / Q000648:ultrastructure*; D013059:Spectrum Analysis, Raman
D010455:Peptides; D012261:Ribonucleoproteins; D011061:Poly A; C015462:polyarginine