Abstract
1H NMR spectroscopy
is used to study the mechanism by which apomyoglobin interacts with heme
to form the holoprotein. In contrast to classic optical studies of the
refolding process by which the native state is reached in a millisecond
time scale, the present results indicate the presence of an intermediate
which converts the holoprotein with a half-life of 5 x 104 s
at physiol. pH. Moreover, .apprx.10% of the intermediate is present at
equil., indicating that the native protein is itself heterogeneous. The
slow rate of conversion suggests that in the native protein either a conformational
uniqueness is controlled by enzyme mediation in the biosynthesis pathway
or heterogeneous heme order is prevalent in vivo.