The formation and the effects of
laser irradiation of the complex formed by protoporphyrin IX (
PPIX) and
tubulin was investigated. We have used
tubulin as a model
protein to investigate whether docked photoactive
ligands can affect the structure and function of
polypeptides upon exposure to visible light. We observed that
laser irradiation in the Soret band prompts bleaching of the
PPIX, which is accompanied by a sharp decrease in the intensity and average fluorescence lifetime of the
protein (dominated by the four
tryptophan residues of the
tubulin monomer). The kinetics indicate non-trivial effects and suggest that the
photosensitization of the
PPIX bound to
tubulin prompts structural alterations of the
protein. These modifications were also observed through changes in the energy transfer between Trp residues and
PPIX. The results suggest that
laser irradiation produces localized partial unfolding of
tubulin and that the changes prompt modification of the formation of microtubules in vitro. Measurements of
singlet oxygen formation were inconclusive in determining whether the changes are prompted by
reactive oxygen species or other excited state mechanisms.