Abstract:
Bacillus thuringiensis Cry4Ba insecticidal crystal protein insertion into lipid
monolayer was investigated by Langmuir-Blodgett technique. Three types of
synthetic lipid, dipalmitoylphosphatidylcholine (DPPC), dioleylethanolamine (DOPE)
and cholesterol (Chol), were used as the model monolayer. The insertion study was
done by compression isotherm where the adsorption kinetics of pure protein, pure
lipid and mix lipid-protein were carried out at the gas/water interface. Protein
molecules only partially inserted some part into the lipid monolayer. This occurred
along with rearrangement of the protein structure at the interface. In lipid layer at π=0,
the amount of adsorbed protein was found to depend on the induction time. There was
an increased area due to protein insertion at all temperatures in all three layers. The
increased area was higher in DPPC and Chol layer than DOPE layer. The inserted
protein alters the elasticity property of the lipid layer as evidenced by a decreased in
the compressibility modulus. The insertion kinetics can be fitted with double
exponential process where the increasing surface pressure (Δπ) from the protein
insertion is fitted as a function of time. Fluorescence spectroscopy revealed that there
is a structural change associated with proteins at the interface.