Chaperone-like activity and surface hydrophobicity of 70S ribosome.
Ribosomes have been shown to mediate refolding of proteins in vitro. In order to understand the mechanism of action, we have explored the 70S ribosome surface for hydrophobicity, one of the important aspects in chaperone-target protein interaction. We find that the 70S ribosome displays significant hydrophobicity on its surface when probed with the hydrophobic fluorophore 8-anilino-1-naphthalene sulfonate. To understand the functional significance of this hydrophobicity we investigated the ability of the ribosome to prevent aggregation of insulin B chain and alpha-lactalbumin induced by reducing the interchain and intrachain disulfide bond respectively with dithiothreitol (DTT) and photo aggregation of gamma-crystallin at 37 degrees C. The 70S ribosome offers complete protection towards light-induced aggregation of gamma-crystallin (at 1:2 (w/w) ratio of crystallin:ribosome) and DTT-induced aggregation of alpha-lactalbumin (at 1:3) and there is appreciable protection (at 1:3) against the aggregation of insulin B chain. We also investigated the role of 70S ribosome in refolding of bovine carbonic anhydrase. Ribosomes improved the folding yield in a concentration-dependent manner. These results clearly demonstrate a general chaperone-like activity of 70S ribosome and implicate its surface hydrophobicity.
Singh
Ranvir
R
Centre for Cellular and Molecular Biology, Uppal Road, 500 007, Hyderabad, India.
Rao
Ch Mohan
ChM
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D004229:Dithiothreitol / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D007328:Insulin / Q000737:chemistry; D007768:Lactalbumin / Q000737:chemistry; D018832:Molecular Chaperones / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism*; D038222:gamma-Crystallins / Q000737:chemistry
D000817:Anilino Naphthalenesulfonates; D004220:Disulfides; D007328:Insulin; D018832:Molecular Chaperones; D038222:gamma-Crystallins; C027132:1-anilino-8-naphthalenesulfonate; D007768:Lactalbumin; D002256:Carbonic Anhydrases; D004229:Dithiothreitol