A structure of a target system (in this study, an actin filament) is likely to be
known from experiment, and as such is in a relatively deep local free-energy
minimum. Hence, the observed values for a CV (Q) are likely to
be normally distributed around a single value (with a roughly harmonic potential
of mean force F(Q)). When a sub-structure such
as an actin monomer is removed to solution, the starting structure (A) will
likely still be near a local free energy minimum, however there may be
alternative lower free-energy configurations (B). The initially-estimated
Lagrange multipliers needed to have the subsystem stay in state A will depend on
whether the system starts in state A or B.