Thermal and GdmCl-induced unfolding transitions of aldolase from Staphylococcus aureus are reversible under a variety of solvent conditions. Analysis of the transitions reveals that no partially folded intermediates can be detected under equilibrium conditions. The stability of the enzyme is very low with a delta G0 value of -9 +/- 2 kJ/mol at 20 degrees C. The kinetics of unfolding and refolding of aldolase are complex and comprise at least one fast and two slow reactions. This complexity arises from prolyl isomerization reactions in the unfolded chain, which are kinetically coupled to the actual folding reaction. Comparison with model calculations shows that at least two prolyl peptide bonds give rise to the observed slow folding reactions of aldolase and that all of the involved bonds are presumably in the trans conformation in the native state. The rate constant of the actual folding reaction is fast with a relaxation time of about 15 s at the midpoint of the folding transition at 15 degrees C. The data presented on the folding and stability of aldolase are comparable to the properties of much smaller proteins. This might be connected with the simple and highly repetitive tertiary structure pattern of the enzyme, which belongs to the group of alpha/beta barrel proteins. Study holds ProTherm entries: 10588, 10589, 10590 Extra Details: alpha/beta barrel; folding intermediate; prolyl isomerization;,protein folding kinetics; two-state model
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:40 p.m.
|Number of data points||7|
|Proteins||Dihydroneopterin aldolase ; Dihydroneopterin aldolase|
|Assays/Quantities/Protocols||Experimental Assay: dHcal ; Experimental Assay: Tm prot_conc:29 micro M ; Experimental Assay: dHvH prot_conc:29 micro M ; Experimental Assay: Tm prot_conc:0.84 micro M ; Experimental Assay: dHvH prot_conc:0.84 micro M ; Experimental Assay: Tm prot_conc:0.84 micro M ; Experimental Assay: dHvH prot_conc:0.84 micro M|
|Libraries||Mutations for sequence MQDTIFLKGMRFYGYHGALSAENEIGQIFKVDVTLKVDLSEAGRTDNVIDTVHYGEVFEEVKSIMEGKAVNLLEHLAERIANRINSQYNRVMETKVRITKENPPIPGHYDGVGIEIVRENK|