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Substitution of non-catalytic stem and loop regions of hammerhead ribozyme with DNA counterparts only increases KM without sacrificing the catalytic step (kcat) : a way to improve substrate-specificity

Shimayama, T.; Sawata, S.; Komiyama, M.; Takagi, Y.; Tanaka, Y.; Wada, A.; Sugimoto, N.; Rossi, J.J.; Nishikawa, F.; Nishikawa, S.

Nucleic Acids Symposium Series 27: 17-18

1992

ISSN/ISBN: 0261-3166
PMID: 1283905
Document Number: 394321
In elucidating structure-function relationships and stabilizing ribozymes in vivo, several chimeric RNA/DNA ribozymes and substrates were chemically synthesized. Measurements of kinetic parameters revealed that the maximally deoxyribonucleotide-substituted ribozyme (DRDRD32) gained the highest catalytic activity reaching the kcat value of > 10 min-1, the highest value ever reported for hammerhead-type ribozymes. Since these chimeric ribozymes are more stable than the wild-type all-RNA ribozymes in vivo and they also possess higher substrate-specificity, they are considered to be better candidates for antiviral therapeutic agents.

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