Previously, we showed that mutants of Thermus thermophilus 3-isopropylmalate dehydrogenase (IPMDH) each containing a residue (ancestral residue) that had been predicted to exist in a postulated common ancestor protein often have greater thermal stabilities than does the contemporary wild-type enzyme. In this study, the combined effects of multiple ancestral residues were analyzed. Two mutants, containing multiple mutations, Sup3mut (Val181Thr/Pro324Thr/Ala335Glu) and Sup4mut (Leu134Asn/Val181Thr/Pro324Thr/Ala335Glu) were constructed and show greater thermal stabilities than the wild-type and single-point mutant IPMDHs do. Most of the mutants have similar or improved catalytic efficiencies at 70 degrees C when compared with the wild-type IPMDH. Study holds ProTherm entries: 22914, 22915, 22916, 22917 Extra Details: 0.5 mM EDTA was added in the experiment Ancestral residue; Protein stability; 3-Isopropylmalate dehydrogenase; Thermus thermophilus
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:54 p.m.
|Number of data points||8|
|Proteins||3-isopropylmalate dehydrogenase ; 3-isopropylmalate dehydrogenase|
|Assays/Quantities/Protocols||Experimental Assay: Tm details:Additives , prot_conc:5.4 microM, buffers:phosphate: 20 mM ; Experimental Assay: Tm prot_conc:10.9 microM, buffers:potassium phosphate: 20 mM, details:Additives EDTA (0.5 mM), ; Derived Quantity: dTm details:Additives , prot_conc:5.4 microM, buffers:phosphate: 20 mM ; Derived Quantity: dTm prot_conc:10.9 microM, buffers:potassium phosphate: 20 mM, details:Additives EDTA (0.5 mM),|
|Libraries||Mutations for sequence MKVAVLPGDGIGPEVTEAALKVLRALDEAEGLGLAYEVFPFGGAAIDAFGEPFPEPTRKGVEEAEAVLLGSVGGPKWDGLPRKIRPETGLLSLRKSQDLFANLRPAKVFPGLERLSPLKEEIARGVDVLIVRELTGGIYFGEPRGMSEAEAWNTERYSKPEVERVARVAFEAARKRRKHVVSVDKANVLEVGEFWRKTVEEVGRGYPDVALEHQYVDAMAMHLVRSPARFDVVVTGNIFGDILSDLASVLPGSLGLLPSASLGRGTPVFEPVHGSAPDIAGKGIANPTAAILSAAMMLEHAFGLVELARKVEDAVAKALLETPPPDLGGSAGTEAFTATVLRHLA|