Stability properties of Aplysia oxymyoglobin: effect of esterification of the heme propionates

Abstract

When compared with sperm whale MbO2 as a reference Aplysia MbO2 is extremely susceptible to autoxidation, and its pH dependence for the reaction is also unusual. Kinetic and thermodynamic analyses have shown that two types of carboxyl group with pKa = 4.3 and 6.1 at 25 degrees C are involved in the stability property of Aplysia MbO2, the protonation of these groups being responsible for an increase in its autoxidation rate in the acidic pH range. Since protoheme has two carboxyl groups of the propionic acid side-chains, we have prepared Aplysia myoglobin containing the dimethylester of protohemin. The autoxidation of this derivative was found to be described by the involvement of only one type of carboxyl group with pKa = 4.4. A possible candidate for the protein residue was therefore discussed on the basis of the known amino acid sequences of three Aplysia myoglobins: it is suggested that Glu-94, next to the proximal His-95, is the most likely candidate.