Jianfeng He

Understanding the folding mechanisms of multi-domain proteins is crucial for gaining insights into protein folding dynamics. The BphC enzyme, a key player in the degradation of polychlorinated biphenyls consists of eight identical subunits, each containing two domains, with each domain comprising two “βαβββ” motifs. In this study, we employed high-temperature molecular dynamics simulations to systematically analyze the unfolding dynamics of a BphC subunit. Our results reveal that the unfolding process of BphC is a complex, multi-intermediate, and multi-phased event. Notably, we identified a thermodynamically stable partially unfolded intermediate. The unfolding sequences, pathways, and rates of the motifs differ significantly. Motif D unfolds first and most rapidly, while Motif C initiates unfolding before Motifs A and B but completes it slightly later. The unfolding behavior of the motifs strongly influences the domain unfolding, leading to the early initiation of Domain 2 unfolding compared to Domain 1, although at a slower rate. The motifs and domains exhibit both independence and cooperativity during the unfolding process, which we interpret through proposed cascading effects. We hypothesize that the folding mechanism of BphC begins with local folding, which propagates through cooperative interactions across structural hierarchies to achieve the folded state. These findings provide new insights into the folding and unfolding mechanisms of multi-domain proteins.

The study of epitopes has a broad range of applications in drug discovery, vaccine design, and immunotherapy. In this study, an epitope prediction method was developed based on the dynamic structure of protein antigens. Solvent accessible surface area, charge, and root mean square fluctuation were introduced as the key residue property parameters. The epitope prediction algorithm was established by constructing a three-parameter complex metrics of seven-peptide groups. The method was applied to predict the epitopes of Exendin-4, an effective antidiabetic drug. The epitopes of both the natural and C-terminal amidated forms of Exendin-4 were predicted and compared in their folded and intermediate states. In the folded state, the epitopes of natural Exendin-4 (His1-Phe6 and Asp9-Val19) were found to be nearly identical to the epitopes of C-terminal aminated Exendin-4 (His1-Thr7 and Asp9-Val19). In the intermediate state, however, the epitopes of natural Exendin-4 (His1-Gly4, Phe6 and Lys12-Arg20) covered fewer amino acids than the epitopes of C-terminal aminated Exendin-4 (His1-Gly4, Phe6, Asp9-Val19 and Trp25-Lys27). The comparison with the results from other prediction tools demonstrates the reliability of our predicted epitopes of Exendin-4.

Proteins are important biological molecules whose structures are closely related to their specific functions. Understanding how the protein folds under physical principles, known as the protein folding problem, is one of the main tasks in modern biophysics. Coarse-grained methods play an increasingly important role in the simulation of protein folding, especially for large proteins. In recent years, we proposed a novel coarse-grained method derived from the topological soliton model, in terms of the backbone C _α chain. In this review, we will first systematically address the theoretical method of topological soliton. Then some successful applications will be displayed, including the thermodynamics simulation of protein folding, the property analysis of dynamic conformations, and the multi-scale simulation scheme. Finally, we will give a perspective on the development and application of topological soliton.

A protein is traditionally visualised as a piecewise linear discrete curve, and its geometry is conventionally characterised by the extrinsically determined Ramachandran angles. However, a protein backbone has also two independent intrinsic geometric structures, due to the peptide planes and the side chains. Here we adapt and develop modern 3D virtual reality techniques to scrutinize the atomic geometry along a protein backbone, in the vicinity of a peptide plane. For this we compare backbone geometry-based (extrinsic) and structure-based (intrinsic) coordinate systems, and as an example we inspect the trans and cis peptide planes. We reveal systematics in the way how a cis peptide plane deforms the neighbouring atomic geometry, and we develop a virtual reality based visual methodology that can identify the presence of a cis peptide plane from the arrangement of atoms in its vicinity. Our approach can easily detect exceptionally placed atoms in crystallographic structures. Thus it can be employed as a powerful visual refinement tool which is applicable also in the case when resolution of the protein structure is limited and whenever refinement is needed. As concrete examples we identify a number of crystallographic protein structures in Protein Data Bank (PDB) that display exceptional atomic positions around their cis peptide planes.

We propose to combine a mean field approach with all atom molecular dynamics into a multistage algorithm that can model protein folding and dynamics over very long time periods yet with atomic level precision. As an example we investigate an isolated monomeric Myc oncoprotein that has been implicated in carcinomas including those in colon, breast and lungs. Under physiological conditions a monomeric Myc is presumed to be an example of intrinsically disordered proteins, that pose a serious challenge to existing modelling techniques. We argue that a room temperature monomeric Myc is in a dynamical state, it oscillates between different conformations that we identify. For this we adopt the C-alpha backbone of Myc in a crystallographic heteromer as an initial Ansatz for the monomeric structure. We construct a multisoliton of the pertinent Landau free energy, to describe the C-alpha profile with ultra high precision. We use Glauber dynamics to resolve how the multisoliton responds to repeated increases and decreases in ambient temperature. We confirm that the initial structure is unstable in isolation. We reveal a highly degenerate ground state landscape, an attractive set towards which Glauber dynamics converges in the limit of vanishing ambient temperature. We analyse the thermal stability of this Glauber attractor using room temperature molecular dynamics. We identify and scrutinise a particularly stable subset in which the two helical segments of the original multisoliton align in parallel, next to each other. During the MD time evolution of a representative structure from this subset, we observe intermittent quasiparticle oscillations along the C-terminal alpha-helix, some of which resemble a translating Davydov's Amide-I soliton. We propose that the presence of oscillatory motion is in line with the expected intrinsically disordered character of Myc.