Effect of polyanion on the acidic conformational transition of native and denatured ferricytochrome c. Circular dichroism study.
Interaction of polyanion poly(vinylsulfate) with oxidized cytochrome c (cyt c) significantly affects the protein main characteristics. One of them, pKa value of acidic transition, was shifted from an apparent pKa value 2.5 (typical for cyt c in low ionic strength solvent) to approximately 5.20 +/- 0.15 upon polyanion binding to the protein, pointing to a likely involvement of histidines 26 and/or 33 in the protein acidic transition in complex with the polyanion. The acidic transition followed at 6 different wavelengths all over circular dichroism spectrum, monitoring different parts of the protein structure, revealed basically two-state character process. Only ellipticity at 262 nm indicated a low-cooperative pH-induced conformational transition in heme region with an apparent pKa approximately 4.34 +/- 0.25 in accordance with absorbance change at 620 nm. Polyanion also interacts with chemically-denatured (in the presence of 9 mol/l urea) state of the protein as it follows from stabilization of protein residual structure at acidic pH and its effect on pKa value of acidic transition of chemically-denatured cyt c. Destabilization effect of polyanions on native and, on the other hand, stabilization influence on partially unfolded conformations of the protein are discussed with an implication for their chaperone-like properties in vivo and in vitro.
Sedlák
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Department of Biochemistry, Faculty of Science, P. J. Safárik University, Kosice, Slovakia. sedlak_er@saske.sk
Antalík
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D000818:Animals; D002942:Circular Dichroism / Q000379:methods*; D003574:Cytochrome c Group / Q000737:chemistry*; D004795:Enzyme Stability; D006736:Horses; D006863:Hydrogen-Ion Concentration; D000071228:Polyelectrolytes; D011108:Polymers / Q000737:chemistry; D011145:Polyvinyls / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D012680:Sensitivity and Specificity
D003574:Cytochrome c Group; D000071228:Polyelectrolytes; D011108:Polymers; D011145:Polyvinyls; C019050:polyvinyl sulfate