Inequivalent conformational response of Chironomus hemoglobins to ligation with O2 and CO. A circular-dichrosim and infrared-spectroscopic study.

For the monomeric hemoglobins III and IV and the dimeric hemoglobins of Chironomus the tryptophan circular dichrosim reports different conformational response to binding of different ligands. The fine structure bands at 292 nm are most strongly developed in the unliganded state and considerably reduced in the oxy form. The spectrum of the CO derivative is intermediate but clearly more deoxy-like. This finding correlates well with the corresponding Bohr effect magnitudes determined by measuring proton release upon ligation. Infrared difference spectroscopy on O2 versus CO derivatives in aqueous solution shows a normal O2 stretching band position at 1107 cm-1 characteristic of asymmetric oxygen binding. The CO stretching band, however, is consistently blue-shifted by 11--13 cm-1 from the 1951 cm-1 position observed with mammalian hemoglobins, indicating reduced CO binding strength. Structural factors relevant to an explanation of the observed phenomena are discussed.