In the case of a probe attached
covalently to a protein, via a linkage through an amine or sulfhydryl groups
for example, or in the case of tryptophan or tyrosine sidechains, considerable
“local” motion of the fluorophore can occur.
In addition, the protein may consist of flexible domains which can
rotate independent of the overall “global” protein rotation. This type of mobility hierarchy is
illustrated on the right for the case of a probe covalently attached to a
dimeric protein
(a) overall L7/L12
rotation
(b) movement of
one C-domain relative to other domains
(c)
movement of dye molecule around its point of attachment