Oligonucleotide-directed mutagenesis has been used to replace alpha-helical glycines in the N-terminal domain of lambda repressor with alanines. Since alanine is a significantly better helix-forming residue than glycine, these changes were predicted to have a stabilizing effect. We show that the Gly46----Ala substitution, the Gly48----Ala substitution, and the double substitution increase the melting temperature of the N-terminal domain by 3-6 degrees. Study holds ProTherm entries: 790, 791, 792, 793, 2273, 2274, 13412, 13413, 13414, 13985, 13986 Extra Details: alpha helix; lambda repressor; stabilizing effect;,melting temperature
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:15 p.m.
|Number of data points||21|
|Proteins||Repressor protein cI ; Repressor protein cI|
|Assays/Quantities/Protocols||Experimental Assay: ddG ; Experimental Assay: dHcal ; Experimental Assay: Tm ; Derived Quantity: dTm|
|Libraries||Mutations for sequence STKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVS|