Effects of temperature, aurintricarboxylic acid and cibacron blue on 2',5'-oligoadenylate binding protein (RNase L) activity in rabbit reticulocyte lysates.
The binding of p3A4,3'-32P [pCp] to rabbit reticulocyte RNase L can be displaced by the trimer and tetramer triphosphates of 2',5'-oligoadenylates (2-5A). Using assay conditions of protein synthesis, 2-5A trimer or tetramer triphosphates are shown to be equally effective when the displacement is done at 4 degrees C (on ice). In contrast, at 30 degrees C, the tetramer triphosphates still displace whereas the trimer triphosphates become ineffective. When lysates are preincubated at temperature ranging from 4 degrees-37 degrees C, the same results are obtained even when the subsequent displacement is done on ice. Incubation temperature also significantly affects the ability of metabolically stable dyes cibacron blue and aurintricarboxylic acid to inhibit RNase L binding activity. Taken together, these results suggest that rabbit reticulocyte RNase L may assume multiple conformations which are differentially affected by various forms of 2-5A or other compounds.
D000818:Animals; D001312:Aurintricarboxylic Acid / Q000494:pharmacology*; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology*; D004722:Endoribonucleases / Q000378:metabolism*; D066298:In Vitro Techniques; D011485:Protein Binding; D011487:Protein Conformation; D011817:Rabbits; D012156:Reticulocytes / Q000187:drug effects / Q000201:enzymology*; D013696:Temperature; D014227:Triazines / Q000494:pharmacology*
D003509:Cyclohexanecarboxylic Acids; D014227:Triazines; D001312:Aurintricarboxylic Acid; C012842:Cibacron Blue F 3GA; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease