Vibrational energy transfer (VET) of proteins at cell membrane plays critical roles in controlling the protein functionalities, but its detection is very challenging. By using a surface-sensitive femtosecond time-resolved sum-frequency generation vibrational spectroscopy with infrared pump, the detection of the ultrafast VET in proteins at cell membrane has finally become possible. The vibrational relaxation time of the N-H groups is determined to be 1.70(±0.05) ps for the α-helix located in the hydrophobic core of the lipid bilayer and 0.9(±0.05) ps for the membrane-bound β-sheet structure. The N-H groups with strong hydrogen bonding gain faster relaxation time. By pumping the amide A band and probing amide I band, the vibrational relaxation from N-H mode to C=O mode through two pathways (direct coupling and through intermediate states) is revealed. The ratio of the pathways depends on the NH⋅⋅⋅O=C hydrogen-bonding strength. Strong hydrogen bonding favors the coupling through intermediate states.
Accurate determination of intermolecular interaction forces at the surface and the interface is essential to identify the nature of interfacial phenomena such as absorption, interfacial assembly, and specific ion effect, but it still represents a major technical challenge. In this study, we proposed a novel method to deduce the interfacial interaction forces by using surface-sensitive second-order Fermi resonant signals, generated in sum frequency generation vibrational spectroscopy (SFG-VS). By investigating the influence of lipid chain length and intermolecular distance on the Fermi resonant signals of phospholipid monolayer at the air/CaF 2 surface and the air/water interface, a linear correlation between the Fermi resonant intensity ratio and the dominated interactions in the lipid monolayer has been observed. It implies that the amplitude of the intensity ratio can be used as an effective in situ vibrational optical ruler to characterize the total intermolecular interaction forces at the surface and the interface. Such a relationship further enables us to elucidate the specific ion effects on the interfacial interactions, allowing us to identify different contributions from van der Waals, electrostatic, and hydration interactions. This study clearly demonstrates the power of the second-order Fermi resonant signals for evaluating the interfacial interaction forces in situ and in real time.
Vibrational energy transfer (VET) of proteins at cell membrane plays critical roles in controlling the protein functionalities, but its detection is very challenging. By using a surface‐sensitive femtosecond time‐resolved sum‐frequency generation vibrational spectroscopy with infrared pump, the detection of the ultrafast VET in proteins at cell membrane has finally become possible. The vibrational relaxation time of the N−H groups is determined to be 1.70(±0.05) ps for the α‐helix located in the hydrophobic core of the lipid bilayer and 0.9(±0.05) ps for the membrane‐bound β‐sheet structure. The N−H groups with strong hydrogen bonding gain faster relaxation time. By pumping the amide A band and probing amide I band, the vibrational relaxation from N−H mode to C=O mode through two pathways (direct coupling and through intermediate states) is revealed. The ratio of the pathways depends on the NH⋅⋅⋅O=C hydrogen‐bonding strength. Strong hydrogen bonding favors the coupling through intermediate states.
The balance of lipid-peptide and peptide-peptide interactions at cell membrane is essential to a large variety of cellular processes. In this study, we have experimentally demonstrated for the first time that sum frequency generation vibrational spectroscopy can be used to probe the peptide-peptide and lipid-peptide interactions in cell membrane in situ and in real time by determination of the line width of amide I band of protein backbone. Using a "benchmark" model of α-helical WALP23, it is found that the dominated lipid-peptide interaction causes a narrow line width of the amide I band, whereas the peptide-peptide interaction can markedly broaden the line width. When WALP23 molecules insert into the lipid bilayer, a quite narrow line width of the amide I band is observed because of the lipid-peptide interaction. In contrast, when the peptide lies down on the bilayer surface, the line width of amide I band becomes very broad owing to the peptide-peptide interaction. In terms of the real-time change in the line width, the transition from peptide-peptide interaction to lipid-peptide interaction is monitored during the insertion of WALP23 into 1,2-dipalmitoyl- sn-glycero-3-phospho-(1'- rac-glycerol) (DPPG) lipid bilayer. The dephasing time of a pure α-helical WALP23 in 1-palmitoyl-2-oleoyl- sn-glycero-3-phospho-(1'- rac-glycerol) and DPPG bilayer is determined to be 2.2 and 0.64 ps, respectively. The peptide-peptide interaction can largely accelerate the dephasing time.
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