Author: Bouvette, Jonathan; Korkut, Dursun Nizam; Fouillen, Aurélien; Amellah, Soumiya; Nanci, Antonio; Durocher, Yves; Omichinski, James G.; Legault, Pascale
Title: High-yield production of human Dicer by transfection of human HEK293-EBNA1 cells grown in suspension Document date: 2018_12_6
ID: 012ipcdr_21
Snippet: In binding studies, we determined K d values of 5 ± 1 nM (WT Dicer) and 9 ± 1 nM (D1320A/D1709A Dicer) between Dicer and pre-let-7a-1. These values are similar to previously reported K d values of 1.8-39 nM obtained for the binding of WT Dicer to pre-let-7a-1 under similar conditions [18, 20, 41, 42] . Moreover, our studies confirm observations with other RNAseIII enzymes that the amino acid substitutions that disrupt magnesium binding in the R.....
Document: In binding studies, we determined K d values of 5 ± 1 nM (WT Dicer) and 9 ± 1 nM (D1320A/D1709A Dicer) between Dicer and pre-let-7a-1. These values are similar to previously reported K d values of 1.8-39 nM obtained for the binding of WT Dicer to pre-let-7a-1 under similar conditions [18, 20, 41, 42] . Moreover, our studies confirm observations with other RNAseIII enzymes that the amino acid substitutions that disrupt magnesium binding in the RNaseIIIa and RNAseIIIb domains do not affect substrate binding to Dicer [43] .
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