Characterization of the polyanion-induced molten globule-like state of cytochrome c.
Cytochrome c (cyt c) undergoes a poly(vinylsulphate) (PVS)-induced transition at slightly acidic pH into a molten globule-like state that resembles the effect that negatively charged membrane surfaces have on this protein. In this work, the thermodynamic properties of the molten globule-like state of cyt c in complex with PVS are studied using differential scanning calorimetry, circular dichroism, fluorescence, and absorbance spectroscopy. The temperature-induced transition of the molten globule-like state of cyt c in the complex with PVS is characterized by a significantly lower calorimetric enthalpy than in the "typical" molten globule state of cyt c, i.e. free protein at pH 2.0 in high ionic strength. Moreover, the thermally-denatured state of cyt c in the complex at pH < 6 contains nearly 50% of the native secondary structure. The dependence of the transition temperature on the pH indicates a role for histidine residues in the destabilization of the cyt c structure in the PVS complex and in stabilization of the denatured state with the residual secondary structure. A comparison of the effects of small anions and polyanions demonstrates the importance of cooperativity among the anions in the destabilization of cyt c. Predictably, other hydrophilic flexible polyanions such as heparin, polyglutamate, and polyadenylate also have a destabilizing effect on the structure of cyt c. However, a correlation between the properties of the polyanions and their effect on the protein stability is still unclear.
Sedlák
Erik
E
Department of Biochemistry, Faculty of Science UPJS, Moyzesova 11, 040 01 Kosice, Slovakia. erik.sedlak@upjs.sk
D000818:Animals; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D045304:Cytochromes c / Q000737:chemistry* / Q000145:classification; D004795:Enzyme Stability; D006736:Horses; D006863:Hydrogen-Ion Concentration; D000071228:Polyelectrolytes; D011108:Polymers / Q000737:chemistry*; D017510:Protein Folding; D013053:Spectrophotometry
D045304:Cytochromes c; D000071228:Polyelectrolytes; D011108:Polymers