4-Aminopyrazolo[3,4-d]pyrimidine (4-APP) as a novel inhibitor of the RNA and DNA depurination induced by Shiga toxin 1.
Shiga toxin 1 (Stx1) catalyses the removal of a unique and specific adenine from 28S RNA in ribosomes (RNA-N-glycosidase activity) and the release of multiple adenines from DNA (DNA glycosylase activity). Added adenine behaves as an uncompetitive inhibitor of the RNA-N-glycosidase reaction binding more tightly to the Stx1-ribosome complex than to the free enzyme. Several purine derivatives and analogues have now been assayed as inhibitors of Stx1. Most of the compounds showed only minor differences in the rank order of activity on the two enzymatic reactions catalysed by Stx1. The survey highlights the importance of the amino group in the 6-position of the pyrimidine ring of adenine. Shifting (2-aminopurine) or substituting (hypoxanthine, 6-mercapto-purine, 6-methylpurine) the group greatly decreases the inhibitory power. The presence of a second ring, besides the pyrimidine one, is strictly required. Substitution, by introducing an additional nitrogen, of the imidazole ring of adenine with triazole leads to loss of inhibitory power, while rearrangement of the nitrogen atoms of the ring from the imidazole to the pyrazole configuration greatly enhances the inhibitory power. Thus 4-aminopyrazolo[3,4-d]pyrimidine (4-APP), the isomer of adenine with the five-membered ring in the pyrazole configuration, is by far the most potent inhibitor of both enzymatic reactions catalysed by Stx1. This finding opens perspectives on therapeutic strategies to protect endothelial renal cells once endocytosis of Stx1 has occurred (haemolytic uraemic syndrome). In the RNA-N-glycosidase reaction 4-APP binds, as adenine, predominantly to the Stx1-ribosome complex (uncompetitive inhibition), while inhibition of the DNA glycosylase activity by both inhibitors is of the mixed type.
Brigotti
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Dipartimento di Patologia Sperimentale dell'Università degli Studi di Bologna, Via San Giacomo 14, I-40126 Bologna, Italy.
Carnicelli
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D
Accorsi
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Rizzi
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Montanaro
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L
Sperti
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D000225:Adenine / Q000494:pharmacology; D000345:Affinity Labels; D001386:Azides / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D003603:Cytotoxins / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism*; D045647:DNA Glycosylases; D007700:Kinetics; D009699:N-Glycosyl Hydrolases / Q000037:antagonists & inhibitors*; D011687:Purines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D012339:RNA, Ribosomal, 28S / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D054788:Ribosome Inactivating Proteins; D022603:Shiga Toxins
D000345:Affinity Labels; D001386:Azides; D001427:Bacterial Toxins; D003603:Cytotoxins; D011687:Purines; D011743:Pyrimidines; D012339:RNA, Ribosomal, 28S; D011994:Recombinant Proteins; D022603:Shiga Toxins; C065795:4-azidophenylpropionamide; D004247:DNA; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins; D000225:Adenine