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.

Probing the Molecular Structure and Dynamics of Membrane-Bound Proteins during Misfolding Processes by Sum-Frequency Generation Vibrational Spectroscopy.

Protein misfolding and amyloid formation are implicated in the protein dysfunction, but the underlying mechanism remains to be clarified due to the lack of effective tools for detecting the transient intermediates. Sum frequency generation vibrational spectroscopy (SFG-VS) has emerged as a powerful tool for identifying the structure and dynamics of proteins at the interfaces. In this review, we summarize recent SFG-VS studies on the structure and dynamics of membrane-bound proteins during misfolding processes. This paper first introduces the methods for determining the secondary structure of interfacial proteins: combining chiral and achiral spectra of amide A and amide I bands and combining amide I, amide II, and amide III spectral features. To demonstrate the ability of SFG-VS in investigating the interfacial protein misfolding and amyloid formation, studies on the interactions between different peptides/proteins (islet amyloid polypeptide, amyloid ß, prion protein, fused in sarcoma protein, hen egg-white lysozyme, fusing fusion peptide, class I hydrophobin SC3 and class II hydrophobin HFBI) and surfaces such as lipid membranes are discussed. These molecular-level studies revealed that SFG-VS can provide a unique understanding of the mechanism of interfacial protein misfolding and amyloid formation in real time, in situ and without any exogenous labeling.

Synthesis and discovery of mitochondria-targeting oleanolic acid derivatives for potential PI3K inhibition.

Oleanolic acid and its derivatives have been widely reported for their antitumor activities. Recently, the introduction of a triphenylphosphonium cation moiety has been described to improve the selectivity and cytotoxicity of pentacyclic triterpenoids by targeting the mitochondria of human cancer cells. In this work, a series of novel mitochondria-targeting oleanolic acid derivatives were synthesized and their antitumor activities assessed. The majority of the compounds are more cytotoxicity to cancer cells than normal cells, especially for 6c with IC50 of 0.81 µM in A549 cells, which showed a slight increase compared to doxorubicin (0.97 µM). Mechanism studies demonstrated that 6c induced apoptosis of A549 cells in a dose-dependent manner, and reactive oxygen species production, mitochondrial membrane potential depolarization, and particularly pro-apoptotic proteins upregulated by western blotting experiment may be responsible for the results. Moreover, 6c arrested the cell cycle at G2/M phase and cell migration in A549 cells. Compound 6c had a comparable or somewhat improved activity to the positive control LY294002 in molecular docking studies and in vitro testing, demonstrating that the apoptosis mechanism may involve inhibition of the PI3K-Akt pathway. These results augur well for the use of 6c as a novel triphenylphosphonium-conjugated anticancer agent.