The hammerhead ribozyme catalyzes the deglycosylation of 2'- mercaptocytidine [6]

Journal of the American Chemical Society

Volumn 122, Issue 17, 2000, Pages 4223-4224

  Hamm, Michelle L ^a   Schwans, Jason P ^a   Piccirilli, Joseph A ^a  

^a UNIVERSITY OF CHICAGO   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CYTIDINE DERIVATIVE; RIBOZYME;

CATALYSIS; CATALYST; CHEMICAL MODIFICATION; DEGLYCOSYLATION; ENZYME MECHANISM; LETTER;

EID: 0034600283     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja993027v     Document Type: Letter

References (20)

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5. To maintain the integrity of the mercapto group was not compromised during radiolabeling and purification, we protected it as a 2-pyridyl disulfide (dHH16SSpyr) before these steps. We generated dHH16SH in situ as described in ref 4.
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9. We also performed the entire set of experiments for the HH8 system (see Supporting Information).
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13. This was also true for reactions with 3′-radiolabeled dHH16SH (see Figure C of Supporting Information).
14. Abasic sites in DNA undergo cleavage of the phosphodiester backbone through a β-elimination mechanism. β-elimination occurs because the 2′-H's at abasic sites are acidic in the ring-opened aldehyde form of the sugar. Removal of the 2′-H generates a resonance stabilized enolate anion that can eliminate the 3′-phosphoryl group. Following this elimination, the 4′-H of the newly formed β-enal becomes acidic and may be removed to create a new enolate that can eliminate the 5′-phosphoryl group. See: Mazumder, A.; Gerlt, J. A. Abalson, M. J.; Stubbe, J.; Cunningham, R. P.; Withka, J.; Bolton, P. H. Biochemistry 1991, 30, 1119.
15. N2-like reaction. Under basic conditions, O-phosphorylated derivatives of 2′-mercaptoethanol react in this way. See: Gay, D. C.; Hamer, N. K. J. Chem. Soc. B 1970, 1123.
16. Wecker et al. previously showed that RNA can catalyze attack of a 5′-phosphorothioate at a carbon center. See: Wecker, M.; Smith, D.; Gold, L. RNA 1996, 2, 982.
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