Fermi Resonance of the N-D Stretching Mode Probing the Local Hydrogen-Bonding Environment in Proteins.

Local H-bonding interactions are crucial for proteins to undergo various structural transitions and form different secondary structures. However, identifying slight distinctions in the local H-bonding of proteins is rather challenging. Here, we demonstrate that the Fermi resonance of the N-D stretching mode can provide an effective probe for the localized H-bonding environment of proteins both at the surface/interface and in the bulk. Using sum frequency generation vibrational spectroscopy and infrared spectroscopy, we established a correlation between the Fermi resonance of the N-D mode and protein secondary structures. The H-bond of N-D···C═O splits the N-D modes into two peaks (∼2410 and ∼2470 cm-1). The relative strength ratio (R) between the ∼2410 cm-1 peak and the ∼2470 cm-1 peak is very sensitive to H-bond strength and protein secondary structure. R is less than 1 for α-helical peptides, while R is greater than 1 for ß-sheet peptides. For R < 2.5, both α-helical/loop structures and ß-sheet structures exhibit almost identical Fermi coupling strengths (W = 28 cm-1). For R > 2.5, W decreases from 28 to 14 cm-1 and depends on the aggregation degree of the ß-sheet oligomers or fibrils. The initial local H-bonding status impacts the misfolding dynamics of proteins at the lipid bilayer interface.

Intermolecular Protein-Water Coupling Impedes the Coupling Between the Amide A and Amide I Mode in Interfacial Proteins.

The coupling between different vibrational modes in proteins is essential for chemical dynamics and biological functions and is linked to the propagation of conformational changes and pathways of allosteric communication. However, little is known about the influence of intermolecular protein-H2O coupling on the vibrational coupling between amide A (NH) and amide I (C═O) bands. Here, we investigate the NH/CO coupling strength in various peptides with different secondary structures at the lipid cell membrane/H2O interface using femtosecond time-resolved sum frequency generation vibrational spectroscopy (SFG-VS) in which a femtosecond infrared pump is used to excite the amide A band, and SFG-VS is used to probe transient spectral evolution in the amide A and amide I bands. Our results reveal that the NH/CO coupling strength strongly depends on the bandwidth of the amide I mode and the coupling of proteins with water molecules. A large extent of protein-water coupling significantly reduces the delocalization of the amide I mode along the peptide chain and impedes the NH/CO coupling strength. A large NH/CO coupling strength is found to show a strong correlation with the high energy transfer rate found in the light-harvesting proteins of green sulfur bacteria, which may understand the mechanism of energy transfer through a molecular system and assist in controlling vibrational energy transfer by engineering the molecular structures to achieve high energy transfer efficiency.

Visualizing Water Monomers and Chiral OH-(H2O) Complexes Infiltrated in a Macroscopic Hydrophobic Teflon Matrix.

Insights into the interaction of fluoroalkyl groups with water are crucial to understanding the polar hydrophobicity of fluorinated compounds, such as Teflon. While an ordered hydrophobic-like 2D water layer has been demonstrated to be present on the surface of macroscopically hydrophobic fluorinated polymers, little is known about how the water infiltrates into the Teflon and what is the molecular structure of the water infiltrated into the Teflon. Using highly sensitive femtosecond sum frequency generation vibrational spectroscopy (SFG-VS), we observe for the first time that monomeric H2O and chiral OH-(H2O) complexes are present in macroscopically hydrophobic Teflon. The species are inhomogeneously distributed inside the Teflon matrix and at the Teflon surface. No water clusters or single-file water "wires" are observed in the matrix. SFG free induction decay (SFG-FID) experiments demonstrate that the OH oscillators of physically absorbed molecular water at the surface dephase on the time scale of <230 fs, whereas the water monomers and hydrated hydroxide ions infiltrated in the Teflon matrix dephase much more slowly (680-830 fs), indicating that the embedded monomeric H2O and OH-(H2O) complexes are decoupled from the outer environment. Our findings can well interpret ultrafast water permeation through fluorous nanochannels and the charging mechanism of Teflon, which may tailor the desired applications of organofluorines.

Ordered Water Layer on the Macroscopically Hydrophobic Fluorinated Polymer Surface and Its Ultrafast Vibrational Dynamics.

Hydrophobic-like water monolayers have been predicted at the metal and some polar surfaces by theoretical simulations. However, direct experimental evidence for the presence of this water layer at surfaces, particularly at biomolecule and polymer surfaces, is yet to be validated at room temperature. Here we observe experimentally that an ordered molecular water layer is present at the hydrophobic fluorinated polymer such as polytetrafluoroethylene (PTFE) surface by using sum frequency generation vibrational spectroscopy. The macroscopic hydrophobicity of PTFE surface is actually hydrophilic at the molecular level. The macroscopically hydrophobic character of PTFE is indeed resulting from the hydrophobicity of the ordered two-dimension (2D) water layer, in which cyclic water tetramer structure is found. The water layer at humidity of ≤40% has a vibrational relaxation time of 550 ± 60 fs. The vibrational relaxation time in the frequency range of 3200-3400 cm-1 shows remarkable difference from the interfacial water at the air/H2O interface and the lipid/H2O interface. No discernible frequency dependence of the vibrational relaxation time is observed, indicating the homogeneous dynamics of OH groups in the water layer. These insights into the water layer at the macroscopically hydrophobic surface may contribute to a better understanding of the hydrophobic interaction and interfacial water dynamics.

Acidic Environment Significantly Alters Aggregation Pathway of Human Islet Amyloid Polypeptide at Negative Lipid Membrane.

The misfolding and aggregation of human islet amyloid polypeptide (hIAPP) at cell membrane has a close relationship with the development of type 2 diabetes (T2DM). This aggregation process is susceptible to various physiologically related factors, and systematic studies on condition-mediated hIAPP aggregation are therefore essential for a thorough understanding of the pathology of T2DM. In this study, we combined surface-sensitive amide I and amide II spectral signals from the protein backbone, generated simultaneously in a highly sensitive femtosecond broad-band sum frequency generation vibrational spectroscopy system, to examine the effect of environmental pH on the dynamical structural changes of hIAPP at membrane surface in situ and in real time. Such a combination can directly discriminate the formation of ß-hairpin-like monomer and oligomer/fibril at the membrane surface. It is evident that, in an acidic milieu, hIAPP slows down its conformational evolution and alters its aggregation pathway, leading to the formation of off-pathway oligomers. When matured hIAPP aggregates are exposed to basic subphase, partial conversion from ß-sheet oligomers into ordered ß-sheet fibrillar structures is observed. When exposed to acidic environment, however, hIAPP fibrils partially converse into more loosely patterned ß-sheet oligomeric structures.