title: Identifying disordered regions in proteins by limited proteolysis.
abstract: Limited proteolysis experiments can be successfully used to detect sites of disorder in otherwise folded globular proteins. The approach relies on the fact that the proteolysis of a polypeptide substrate requires its binding in an extended conformation at the protease's active site and thus an enhanced backbone flexibility or local unfolding of the site of proteolytic attack. A striking correlation was found between sites of limited proteolysis and sites of enhanced chain flexibility of the polypeptide chain, this last evaluated by the crystallographically determined B-factor. In numerous cases, it has been shown that limited proteolysis occurs at chain regions characterized by missing electron density and thus being disordered. Therefore, limited proteolysis is a simple and reliable experimental technique that can detect sites of disorder in proteins, thus complementing the results that can be obtained by the use of other physicochemical and computational approaches.
authors: CRIBI Biotechnology Centre, University of Padua, Padua, Italy. angelo.fontana@unipd.it
authors: Angelo
authors: A
authors: Fontana
authors: Patrizia Polverino
authors: PP
authors: de Laureto
authors: Barbara
authors: B
authors: Spolaore
authors: Erica
authors: E
authors: Frare
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000378:metabolism; D020134:Catalytic Domain; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009211:Myoglobin / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D010447:Peptide Hydrolases / Q000737:chemistry / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D059748:Proteolysis
chemical_list: D001059:Apoproteins; D009211:Myoglobin; D010446:Peptide Fragments; D011480:Protease Inhibitors; D011506:Proteins; C010857:apomyoglobin; D010447:Peptide Hydrolases