This research is led by Prof. Wen-Bin Zhang (Faculty of Chemistry and Molecular Engineering, Peking College & Beijing Academy of Synthetic Intelligence) and Dr. Jing Fang (Faculty of Chemistry and Molecular Engineering, Peking College). A single-domain protein catenane refers to 2 mechanically interlocked polypeptide rings that fold synergistically right into a compact and built-in construction, which is extraordinarily uncommon.
This design was achieved by rewiring the connectivity between secondary motifs to introduce synthetic entanglement, and synthesis was readily achieved by a sequence of programmed streamlined post-translational processing occasions in cells with no further in vitro reactions.
The one-domain catenane cat-DHFR was completely characterised. Proof from mixed SDS-PAGE, SEC, LC-MS, IMS-MS, and proteolytic digestion experiments unambiguously proved its topology. The cat-DHFR displays enhanced anti-aggregation properties and has a Tm that’s 6 °C larger than the linear management.
Though the catalytic exercise of cat-DHFR is diminished owing to its decreased affinity towards the substrate and cofactor, it has higher thermal resilience than l-DHFR. Even after incubation at 70 °C for 10 min, cat-DHFR retained over 70% of the catalytic exercise, whereas the linear management misplaced virtually all exercise.
The analysis crew anticipate that this technique might be typically relevant to different single-domain proteins, together with these with folds just like DHFR or with fully different folds. The supply of those single-domain protein catenanes facilitates the elucidation of topological results on construction–property relationships.
The outcomes additional indicate that it’s attainable to map the present linear protein universe into single-domain protein catenanes with well-preserved features and extra advantages, opening up new territory for protein molecules.
Surpassing the linear paradigm of pure protein molecules, these topological proteins are multi-chain, multi-dimensional molecules with useful advantages of topology, wealthy design chance and wonderful evolvability. As a brand new class of protein molecules, they maintain nice potential for a broad variety of functions, together with, however not restricted to, industrial enzymes, antibodies, cytokines, and biomaterials.
