Molecular Insights into the Improved Bioactivity of Interferon Conjugates Attached to a Helical Polyglutamate.

Attaching polymers, especially polyethylene glycol (PEG), to protein drugs has emerged as a successful strategy to prolong circulation time in the bloodstream. The hypothesis is that the flexible chain wobbles on the protein's surface, thus resisting potential nonspecific adsorption. Such a theoretical framework may be challenged when a helical polyglutamate is used to conjugate with target proteins. In this study, we investigated the structure-activity relationships of polyglutamate-interferon conjugates P(EG3Glu)-IFN using molecular simulations. Our results show that the local crowding effect induced by oligoethylene glycols (i.e., EG3) is the primary driving force for helix formation in P(EG3Glu), and its helicity can be effectively increased by reducing the free volume of the two termini. Furthermore, it was found that the steric hindrance induced by IFN is not conductive to the helicity of P(EG3Glu) but contributes to its dominant orientation relative to interferon. The orientation of IFN relative to the helical P(EG3Glu) can help to protect the protein drug from neutralizing antibodies while maintaining its bioactivity. These findings suggest that the helical structure and its orientation are critical factors to consider when updating the theoretical framework for protein-polymer conjugates.

Protecting Enzymatic Activity via Zwitterionic Nanocapsulation for the Removal of Phenol Compound from Wastewater.

Horseradish peroxidase (HRP) holds great potential in wastewater treatment. However, its instability in harsh environments remains a major issue. Various immobilization technologies were developed to retain enzyme stability at the cost of its effectiveness. We demonstrate that zwitterionic encapsulation of HRP retained both protein stability and activity to a large degree. In a water treatment study, encapsulating HRP into a zwitterionic nanogel resulted in a three-fold increase in the catalytic oxidation efficiency of phenol molecules. In addition, zwitterionic nanocapsules exhibited the best performance when compared with nanocapsules made from other hydrophilic polymers. These results indicated that zwitterionic HRP nanocapsules hold great potential in the decontamination of organic pollutants from wastewater.

Inhibition of the fibrillation of highly amyloidogenic human calcitonin by cucurbit[7]uril with improved bioactivity.

Protein/peptide fibrillation is an important challenge for biotechnological drug development. Salmon calcitonin (sCT) is currently used in the clinical treatment of bone-related diseases such as osteoporosis and hypercalcemia, but it still has the risk of immune responses. Although human calcitonin (hCT) would be a better choice in terms of immunogenicity, it has a strong tendency to irreversibly aggregate in aqueous solutions and form long amyloid fibrils, which significantly reduces its bioavailability and therapeutic potency. Here, we demonstrate that cucurbit[7]uril (CB[7]) can inhibit hCT fibrillation by supramolecular interaction with its aromatic groups (affinity: Phe16 > Tyr12 > Phe19 > Phe22). The hCT-CB[7] complex exhibits low cytotoxicity, even promotes osteoblast proliferation and osteogenic capacity of MC3T3 cells. Meanwhile the hCT-CB[7] complexes shows higher bioactivity compared to hCT in reducing blood calcium levels in rats, and also decreases the immunogenicity of hCT. These results suggest that CB[7] has the potential to improve the therapeutic potency of amyloidogenic protein/peptide drugs such as hCT.

Tumor-targeted paclitaxel delivery and enhanced penetration using TAT-decorated liposomes comprising redox-responsive poly(ethylene glycol).

To combine the advantage of poly(ethylene gylcol) (PEG) for longer circulation and cell-penetrating peptides (CPPs) for efficient cellular uptake, paclitaxel (PTX)-loaded liposomes functionalized with TAT, the most frequently used CPP, and cleavable PEG via a redox-responsive disulfide linker (PTX-C-TAT-LP) were successfully developed here. Under physiological conditions, TAT was shielded by PEG layer and liposomes exhibited a long blood circulation. At tumor site, PEG could be detached in the presence of exogenous reducing agent [glutathione (GSH)] and TAT was exposed to facilitate cell internalization. In the presence of GSH, the liposomal vesicle C-TAT-LP showed increased cellular uptake and improved three-dimensional tumor spheroids penetration in vitro compared with analogous stable shielded liposomes. C-TAT-LP achieved enhanced tumor distribution and demonstrated superior delivery efficiency in vivo. PTX-C-TAT-LP with GSH strongly inhibited the proliferation of murine melanoma B16F1 tumor cells in vitro and in vivo with the tumor inhibition rate being 69.4% on B16F1-bearing mice. In addition, the serum aspartate transaminase level, alanine transaminase level, and creatine kinase level were almost completely within normal range in the PTX-C-TAT-LP with GSH group, revealing PTX-C-TAT-LP with GSH had no obvious drug-related adverse events for liver and heart. Taken together, C-TAT-LP is a promising tumor-targeting drug carrier.

Construction of an implicit membrane environment for the lattice Monte Carlo simulation of transmembrane protein.

Due to the complexity of biological membrane, computer simulation of transmembrane protein's folding is challenging. In this paper, an implicit biological membrane environment has been constructed in lattice space, in which the lipid chains and water molecules were represented by the unoccupied lattice sites. The biological membrane was characterized with three features: stronger hydrogen bonding interaction, membrane lateral pressure, and lipophobicity index for the amino acid residues. In addition to the hydrocarbon core spanning region and the water solution, the lipid interface has also been represented in this implicit membrane environment, which was proved to be effective for the transmembrane protein's folding. The associated Monte Carlo simulations have been performed for SARS-CoV E protein and M2 protein segment (residues 18-60) of influenza A virus. It was found that the coil-helix transition of the transmembrane segment occurred earlier than the coil-globule transition of the two terminal domains. The folding process and final orientation of the amphipathic helical block in water solution are obviously influenced by its corresponding hydrophobicity/lipophobicity. Therefore, this implicit membrane environment, though in lattice space, can make an elaborate balance between different driving forces for the membrane protein's folding, thus offering a potential means for the simulation of transmembrane protein oligomers in feasible time.