Graduate Thesis Or Dissertation
 

Solubilization of acid nonextractable flavin-linked protein by cyanide

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  • It has been reported that cyanide inactivates the succinate dehydrogenase activity of HMP toward cytochrome c, oxygen, methylene blue and DCIP. In the present study, it was found that cyanide also solubilizes the SDH from HMP, this is reflected by the extent of solubilization of acid nonextractable flavin in the supernatant after cyanide treatment. The solubilization reaction is dependent on a variety of factors like temperature, cyanide concentration, time of exposure to cyanide and the age of HMP but independent of pH. The soluble SDH was not reactive towards methylene blue, DCIP, and cytochrome c but was active toward PMS and ferricyanide. Incubation of HMP with 40 mM succinate or dithionite completely prevented the solubilization of flavin by cyanide indicating that cyanide acted only on the oxidized form of the enzyme. The activity towards PMS was however very small and hence a differential extraction procedure has been developed to improve the specific activity. This was achieved by a preliminary washing whereby most of the inactive proteins were removed followed by a second extraction resulting in considerable increase in the specific activity.
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