Interaction of native ovalbumin (OVA) with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) Langmuir-Blodgett monolayer has been studied at the air-water interface. A compressibility study shows the positive association with DPPC. Adsorption kinetics shows that the protein adsorption is a one-step process and the amount of protein adsorbed depends on the concentration of protein at the water subphase. Incorporation of protein into the DPPC layer is surface-pressure dependent. The compressibility study indicates that the DPPC-OVA interaction is hydrophobic in nature and structural reorganization is eminent to adjust the hydrophobic mismatch between DPPC acyl chains and OVA hydrophobic moieties. At higher pressure, OVA tends to squeeze out from the DPPC monolayer. A nanometer scale FE-SEM image confirms this observation. Globular aggregates of protein of dimension 60-80 nm were observed in DPPC-OVA supported film. Steady-state fluorescence spectroscopy suggests that the tryptophan residues of OVA are main emitting species. The blue shift of tryptophan fluorescence in supported film may be due to the tryptophan molecule of protein exposed to the hydrophobic air phase.

In this article, we report the results of the extent of interaction as well as the formation of a bioconjugate of human hemoglobin (Hb) with silver (Ag). The complexation process and conformational changes are characterized using different spectroscopic and microscopic techniques. The UV-vis study demonstrates the perturbation of the soret/heme band and generates conformational heterogeneity within the heme group in the presence of silver. A fluorescence study suggests that the Tryptophan (Trp) residues of Hb are in a more polar environment after conjugation. Initial fluorescence enhancement with addition of silver is due to metal-enhanced fluorescence. Moreover, the fluorescence quenching after the formation of the Hb-Ag bioconjugate follows the modified Stern-Volmer (S-V) plot. The S-V plot along with the time-resolved fluorescence study indicates the presence of both static and dynamic types of quenching. In addition, the reduction potential values of the entities (Hb-heme, Ag(+), and Trp) indicate the possible electron transfer. The secondary structure calculation from CD and FTIR spectra indicate alpha-helix to beta-sheet conversion, and unfolding of Hb is also responsible for the bioconjugate formation. In addition, FE-SEM, phase contrast inverted microscopy (PCIM) images demonstrate the formation of the silver-protein bioconjugate. The overall data show that there is a change in the secondary as well as the tertiary structure of Hb after conjugation with silver.

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We report here the effect of salt (KCl) on the interfacial surface activity of yeast alcohol dehydogenease (ADH) at air/water interface using the Langmuir-Blodgett technique. Effect of salt content in the water subphase on ADH structure has been studied. The change of area/molecule, compressibility, rigidity, and unfolding of ADH are insignificant up to 10 mM KCl concentration. The significant changes are observed above 0.1 M KCl concentrations. Observations are explained in the context of DLVO theory. FTIR study of amide band together with AFM imaging of ADH monolayer indicate that KCl perturbs the ADH monolayer by the increment of beta-structure resulting into larger unfolding and intermolecular aggregates at high salt concentration.