Selected article for: "bio layer and real time"
Author: Aaron S. Mendez; Carolin Vogt; Jens Bohne; Britt A. Glaunsinger
Title: Site specific target binding controls RNA cleavage efficiency by the Kaposi’s sarcoma-associated herpesvirus endonuclease SOX
  • Document date: 2018_5_13
    • ID: 298cbr1x_14
    Snippet: To explore the importance of the residues involved in SOX binding and cleavage, we engineered 3 mutants of the LIMD1 54 substrate (Fig. 5A) . First, we preserved the loop structure but replaced the 3 adenosines bound by SOX (residues 39-43) with guanosines (LIMD1 3xA-G). Second, we mutated the residue located at the predicted SOX cut site that was also protected in the footprinting assay (LIMD1 A-G). This mutant has been previously identified to .....
    Document: To explore the importance of the residues involved in SOX binding and cleavage, we engineered 3 mutants of the LIMD1 54 substrate (Fig. 5A) . First, we preserved the loop structure but replaced the 3 adenosines bound by SOX (residues 39-43) with guanosines (LIMD1 3xA-G). Second, we mutated the residue located at the predicted SOX cut site that was also protected in the footprinting assay (LIMD1 A-G). This mutant has been previously identified to block SOX cleavage in vivo (20). Third, we largely The predicted structures of the LIMD1 3XA-G and LIMD1 zipper mutants were verified by in-line probing (Fig. 5, S6 ). Real-time binding kinetics for SOX with WT LIMD1 54 and each of the 3 mutant substrates were then measured using bio-layer interferometry (BLI).

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