title: Effect of deamidation on folding of ribonuclease A.
abstract: The folding of ribonuclease A (RNase A) has been extensively studied by characterizing the disulfide containing intermediates using different experimental conditions and analytical techniques. So far, some aspects still remain unclear such as the role of the loop 65-72 in the folding pathway. We have studied the oxidative folding of a RNase A derivative containing at position 67 the substitution Asn --> isoAsp where the local structure of the loop 65-72 has been modified keeping intact the C65-C72 disulfide bond. By comparing the folding behavior of this mutant to that of the wild-type protein, we found that the deamidation significantly decreases the folding rate and alters the folding pathway of RNase A. Results presented here shed light on the role of the 65-72 region in the folding process of RNase A and also clarifies the effect of the deamidation on the folding/unfolding processes. On a more general ground, this study represents the first characterization of the intermediates produced along the folding of a deamidated protein.
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Salerno, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Orrù
authors.forename: L
authors.initials: L
authors.lastname: Vitagliano
authors.forename: L
authors.initials: L
authors.lastname: Esposito
authors.forename: L
authors.initials: L
authors.lastname: Mazzarella
authors.forename: G
authors.initials: G
authors.lastname: Marino
authors.forename: M
authors.initials: M
authors.lastname: Ruoppolo
mesh_terms: D000577:Amides / Q000378:metabolism / Q000494:pharmacology*; D019943:Amino Acid Substitution; D000818:Animals; D002417:Cattle; D004220:Disulfides; D005978:Glutathione / Q000494:pharmacology; D006860:Hydrogen Bonding; D007700:Kinetics; D010084:Oxidation-Reduction; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics
chemical_list: D000577:Amides; D004220:Disulfides; D012259:Ribonuclease, Pancreatic; D005978:Glutathione