Equilibrium studies of the reaction of turkey (Meleagris gallopavo) haemoglobin sulphydryl groups with 5,5′-Dithiobis(2-nitrobenzoate): tertiary conformational change in turkey haemoglobin induced by inositol hexakisphosphate.

The red blood cell of turkey contains two haemoglobin types, major and mi-nor components. In the present study, the equilibrium constant, Kequ, for the reaction of 5,5′-dithiobis(2-nitrobenzoate), DTNB, with the sulphydryl group of the major turkey aquomethaemoglobin was determined at 25°C as a function of pH. Kequ varies by about 2 to 3 orders of magnitude between pH 5.6 and 9.0 for both haemoglobin [stripped and in the presence of inositol hexakisphosphate (inositol-P6)]. Calculations from the pH dependence of Kequ showed that in the r ⇌ t tertiary conformational transition of aquomethae-moglobin, the t isomer population was 0.26 %. In the presence of inositol-P6, the t isomer population increased to 9.08 %. The results showed that while inositol-P6 increased the relative population of the t tertiary conformation by changing the relative distribution of two protein conformations, it had no effect on Kequ. The effect of Inositol-P6 on the nature and number of groups linked to the DTNB reaction was also determined.