The inhibition of glucosylceramide beta-glucosidase and other acid hydrolases by nucleic acids.
During the isolation of the activator protein for glucosylceramide beta-glucosidase, we found that certain column fractions contained an inhibitor of the enzyme. After separation from the activator protein by a DEAE-Sephacel column, the inhibitor was purified further with a Spehadex G-75 column. The u.v. absorption spectrum of the purified material was similar to that of nucleic acids and the protein content of the purified material was negligible. Furthermore the purified inhibitor reacted with orcinol but not with diphenylamine, and its inhibitory activity was completely destroyed by treatment with RNAases. It seems likely that the purified inhibitor was tRNA. Authentic RNA, tRNA and DNA had similar inhibitory effects on beta-glucosidase (Ki 17 micrograms/ml for tRNA, noncompetitive toward the substrate). The inhibitory effect of nucleic acids was not fully overcome by an excess amount of the activator protein, but phosphatidylserine could restore the activity to normal. Tests with several other hydrolases revealed that the inhibitory effect of nucleic acids was fairly general.
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Mental Health Research Institute, University of Michigan, Ann Arbor 48104.
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D004247:DNA / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D005959:Glucosidases / Q000037:antagonists & inhibitors*; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D006023:Glycoproteins; D006867:Hydrolases / Q000037:antagonists & inhibitors; D007700:Kinetics; D011506:Proteins / Q000494:pharmacology; D012313:RNA / Q000494:pharmacology*; D012343:RNA, Transfer / Q000302:isolation & purification / Q000494:pharmacology; D012260:Ribonucleases; D049231:Saposins
D006023:Glycoproteins; D011506:Proteins; D049231:Saposins; D012313:RNA; D004247:DNA; D012343:RNA, Transfer; D006867:Hydrolases; D012260:Ribonucleases; D005959:Glucosidases; D005962:Glucosylceramidase