Sulfhydryl groups and the interaction between the hemes in hemoglobin

1. Dialysis of a hemoglobin solution against slightly alkaline buffer (pH 8.68) causes a decrease in heme-heme interaction. The value of n in Hill's equation drops from 2.9 to 2.0, while the oxygen affinity increases slightly. Addition of glutathione largely reverses the effects of dialysis (n rises from 2.0 to 2.5 to 2.6). 2. Addition of the sulfhydryl reagent, p-chloromercuribenzoate (1.8 x 10(-3)M), results in a large though not complete blocking of heme-heme interaction. The value of n drops from 2.9 to 1.4. This effect is largely reversed with glutathione (n rises to 2.6).