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In one such temperature-sensitive mutant, a highly conserved asparagine residue in the sensor I motif was changed to alanine.
Lysine 767 within the ARKS motif was changed to arginine.
We generated a mutant in which the consensus motif was changed from VKDE to AKAA (Ago2-3mutCS).
The PxDxL, PxVxS and both IxPxV amino acid motifs were changed to AxAxA, and the LxVxV motif was changed to LxAxA.
Two C. jejuni ciaD mutants were generated; the MAP kinase docking motif was deleted (ΔMKD) in one mutant and the proline-directed phosphorylation motif was changed to an alanine (ΔS/T P) in the other mutant.
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Three p130Cas mutants were expressed: an SD mutant (15F) in which all fifteen YxxP motif tyrosines were changed to phenylalanines, an SBD mutant (mPR) in which prolines in the SH3-binding motif are changed to alanines, and a double mutant (15F/mPR) in which both the SD and SBD are changed.
Instead of the high-scoring CATAAT, the motif is changed to ATTAAT.
The terminal CGTC motif is changed to TGTC or CCTC in many repeats.
The cysteine in the DHHC motif is changed to a serine residue in the D. melanogaster DHHC β1,4-N-acetylgalactosaminyl-transferase B pilot (GABPI).
When the cysteine residues of the radical SAM motif were changed to alanines by site-directed mutagenesis, the protein was found to bind approximately eight irons and sulfides, consistent with the presence of two remaining [4Fe 4S] clusters.
Upon generation of a BtrN variant in which the three cysteines of the radical SAM motif were changed to alanine residues, the enzyme only bound a single [4Fe 4S] cluster based on quantitative elemental analyses and Mössbauer characterization (Table 2).
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