title: Mechanism of induced folding: Both folding before binding and binding before folding can be realized in staphylococcal nuclease mutants.
abstract: A considerable number of functional proteins are unstructured under physiological condition. These "intrinsically disordered" proteins exhibit induced folding when they bind their targets. The induced folding comprises two elementary processes: folding and binding. Two mechanisms are possible for the induced folding: either folding before binding or binding before folding. We found that these two mechanisms can be distinguished by the target-concentration dependence of folding kinetics. We also created two types of mutants of staphylococcal nuclease showing the different inhibitor-concentration dependence of induced folding kinetics. One mutant obeys the scheme of binding before folding, while the other the folding before binding. This is the first experimental evidence demonstrating that both mechanisms are realized for a single protein. Binding before folding is possible, when the protein lacks essential nonlocal interaction to stabilize the native conformation. The results cast light on the protein folding mechanism involved in the intrinsically disordered proteins.
authors.affiliation: Graduate School of Materials Science, Nara Institute of Science and Technology, Ikoma, Nara 630-0192, Japan.
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Onitsuka
authors.forename: Hironari
authors.initials: H
authors.lastname: Kamikubo
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Yamazaki
authors.forename: Mikio
authors.initials: M
authors.lastname: Kataoka
mesh_terms: D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D008024:Ligands; D008836:Micrococcal Nuclease / Q000737:chemistry* / Q000378:metabolism*; D050505:Mutant Proteins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D017510:Protein Folding; D012996:Solutions
chemical_list: D004791:Enzyme Inhibitors; D008024:Ligands; D050505:Mutant Proteins; D012996:Solutions; D008836:Micrococcal Nuclease