Escherichia coli phosphofructokinase-2 (Pfk-2) is a homodimer whose subunits consist of a large domain and an additional beta-sheet that provides the interfacial contacts between the subunits, creating a beta-barrel flattened-like structure with the adjacent subunit's beta-sheet. To determine how the structural organization of Pfk-2 determines its stability, the reversible unfolding of the enzyme was characterized under equilibrium conditions by enzymatic activity, circular dichroism, fluorescence and hydrodynamic measurements. Pfk-2 undergoes a cooperative unfolding/dissociation process with the accumulation of an expanded and unstructured monomeric intermediate with a marginal stability and a large solvent accessibility with respect to the native dimer. Study holds ProTherm entries: 25001, 25002, 25003, 25004 Extra Details: 10 mM DTT was added in the experiment; Native to intermediate Reversible unfolding; Subunit interaction; Folding intermediate; Dimer monomer transition; Ribokinase superfamily; Bimolecular domain
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:55 p.m.
|Number of data points||8|
|Proteins||ATP-dependent 6-phosphofructokinase isozyme 2 ; ATP-dependent 6-phosphofructokinase isozyme 2|
|Assays/Quantities/Protocols||Experimental Assay: m ; Experimental Assay: dG_H2O ; Experimental Assay: m ; Experimental Assay: dG_H2O ; Experimental Assay: m ; Experimental Assay: dG_H2O|
|Libraries||Mutations for sequence MVRIYTLTLAPSLDSATITPQIYPEGKLRCTAPVFEPGGGGINVARAIAHLGGSATAIFPAGGATGEHLVSLLADENVPVATVEAKDWTRQNLHVHVEASGEQYRFVMPGAALNEDEFRQLEEQVLEIESGAILVISGSLPPGVKLEKLTQLISAAQKQGIRCIVDSSGEALSAALAIGNIELVKPNQKELSALVNRELTQPDDVRKAAQEIVNSGKAKRVVVSLGPQGALGVDSENCIQVVPPPVKSQSTVGAGDSMVGAMTLKLAENASLEEMVRFGVAAGSAATLNQGTRLCSHDDTQKIYAYLSR|