Kinetics of cyanide binding as a probe of local stability/flexibility of cytochrome c.
Effect of anions of the Hofmeister series (thiocyanate, perchlorate, iodide, bromide, nitrate, chloride, sulfate, and phosphate) on local and global stability and flexibility of horse heart ferricytochrome c (cyt c) has been studied. Global stability of cyt c was determined by iso/thermal denaturations monitored by change in ellipticity in the far-UV region and its local stability was determined from absorbance changes in the Soret region. Particularly, relative stability/flexibility of the Met80-heme iron bond has been assessed by analysis of binding of cyanide into the heme iron. Both global and local stabilities of cyt c exhibited monotonous increase induced by a change of anion from chaotropic to kosmotropic species. However, this monotonous dependence was not observed for the rate constants of cyanide association with cyt c. As expected more chaotropic ions induced lower stability of protein and faster binding of cyanide but this correlation was reversed for kosmotropic anions. We propose that the unusual bell-shaped dependence of the rate constant of cyanide association is a result of modulation of Met80-heme iron bond strength and/or flexibility of heme region by Hofmeister anions independently on global stability of cyt c. Further, our results demonstrate sensitivity of cyanide binding to local change in stability/flexibility in the heme region of cyt c.
Varhac
Rastislav
R
Department of Biochemistry, Faculty of Science, P.J. Safárik University, 040 01 Kosice, Slovakia.
Tomásková
Natasa
N
Fabián
Marián
M
Sedlák
Erik
E
D000818:Animals; D002942:Circular Dichroism; D003486:Cyanides / Q000737:chemistry*; D045304:Cytochromes c / Q000737:chemistry*; D006418:Heme / Q000737:chemistry; D006736:Horses; D007501:Iron / Q000737:chemistry; D007700:Kinetics; D011485:Protein Binding; D013816:Thermodynamics
D003486:Cyanides; D006418:Heme; D045304:Cytochromes c; D007501:Iron