title: Change of the protein p53 electrochemical signal according to its structural form - quick and sensitive distinguishing of native, denatured, and aggregated form of the "guardian of the genome".
abstract: Presence of mutated and/or structurally modified (e.g., denatured, aggregated) protein p53 form is associated with several disorders such as Alzheimer's disease, Parkinson's disease, prion diseases, and many types of tumours. The aim of this work was to distinguish native, denatured and aggregated form of full-length p53 by flow injection analysis coupled with electrochemical detector (FIA-ED). Firstly FIA-ED method used for protein native form determination was optimized (detection limit 45.8 amol per 5 mul injection; 3 x S/N). In addition the technique was applied to identify p53 structural forms (denatured and aggregated). It was found out that denatured form provides about three times higher electrochemical response (protein structure unfolding, approach of more electroactive centers - aminoacid residues - towards electrode surface) in comparison with native form. On the other hand, aggregated form offers lower response (steric eclipse of electroactive protein parts) when compared with the signal of native form. The obtained data show that we are not only able to sensitively determine native, denatured, and aggregated structural forms of p53 protein but also to distinguish them.
authors.affiliation: Department of Analytical Chemistry, Faculty of Science, Masaryk University, Kotlarska 2, 611 37, Brno, Czech Republic.
authors.forename: David
authors.initials: D
authors.lastname: Potesil
authors.forename: Radka
authors.initials: R
authors.lastname: Mikelova
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
authors.forename: Rene
authors.initials: R
authors.lastname: Kizek
authors.forename: Richard
authors.initials: R
authors.lastname: Prusa
mesh_terms: D004563:Electrochemistry; D017022:Flow Injection Analysis / Q000379:methods*; D006801:Humans; D011487:Protein Conformation; D011489:Protein Denaturation; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry*
chemical_list: D016159:Tumor Suppressor Protein p53