title: A distinct intermediate of RNase A is induced by sodium dodecyl sulfate at its pK(a).
abstract: The chemical denaturation of RNase A was found to be mediated by sodium dodecyl sulfate (SDS) at various pH. The characterization of the unfolding pathway was investigated by spectrophotometry and differential scanning calorimetry (DSC), and was analyzed by multivariate curve resolution (MCR) as a chemometric method. The spectrophotometric titration curve of RNase A upon interaction with SDS indicated a distinct complex intermediate in glycine buffer at pH 3.3. This was accompanied with the catalytic activation of the enzyme and was concurrent with maximum population of the intermediate, determined by MCR. This was confirmed by the DSC profile of RNase A in the presence of SDS, indicated by two transitions in thermal unfolding. The kinetic data on the unfolding process of RNase A upon addition of SDS showed a two-phase pathway under the same conditions. The intermediate appeared at low pH especially at the pK(a) of SDS (pH 3.3). These results provide strong evidence of the influence of low pH (around the pK(a) of SDS) on the existence of an intermediate upon interaction of RNase A with SDS.
authors.affiliation: Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran. moosavi@ut.ac.ir
authors.forename: A A
authors.initials: AA
authors.lastname: Moosavi-Movahedi
authors.forename: M
authors.initials: M
authors.lastname: Gharanfoli
authors.forename: K
authors.initials: K
authors.lastname: Nazari
authors.forename: M
authors.initials: M
authors.lastname: Shamsipur
authors.forename: J
authors.initials: J
authors.lastname: Chamani
authors.forename: B
authors.initials: B
authors.lastname: Hemmateenejad
authors.forename: M
authors.initials: M
authors.lastname: Alavi
authors.forename: A
authors.initials: A
authors.lastname: Shokrollahi
authors.forename: M
authors.initials: M
authors.lastname: Habibi-Rezaei
authors.forename: C
authors.initials: C
authors.lastname: Sorenson
authors.forename: N
authors.initials: N
authors.lastname: Sheibani
mesh_terms: D000465:Algorithms; D000704:Analysis of Variance; D002152:Calorimetry, Differential Scanning; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004790:Enzyme Induction / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D011489:Protein Denaturation; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000096:biosynthesis*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D012967:Sodium Dodecyl Sulfate; D012259:Ribonuclease, Pancreatic