Bioengineering strategies to generate artificial protein complexes

Abstract

For many applications, increasing synergy between distinct proteins through organization is important for the specificity, regulation, and overall reaction efficiency. Although there are many examples of protein complexes in nature, a generalized method to create these complexes remains elusive. Many conventional techniques such as random chemical conjugation, physical adsorption onto surfaces, and encapsulation within matrices are imprecise approaches and can lead to deactivation of protein native functionalities. More "bio-friendly" approaches such as genetically fused proteins and biological scaffolds often can result in low yields and low complex stability. Alternatively, site-specific protein conjugation or ligation can generate artificial protein complexes that preserve the native functionalities of protein domains and maintain stability through covalent bonds. In this review, we describe three distinct methods to synthesize artificial protein complexes (genetic incorPoration of unnatural amino acids to introduce bio-orthogonal azide and alkyne groups to proteins, split-intein based expressed protein ligation, and sortase mediated ligation) and highlight interesting applications for each technique.

Broadening the scope of sortagging

“…Excellent articles summarized the different sortase classes with their functions in nature and structures, 2,4 mechanistic studies, 3,4 sortase inhibitors, 5,6 pili construction, 76,77 peptide and protein cyclisation, 63,78 general application elds and the use for protein engineering, [79][80][81][82][83][84][85] as well as the improvement of SML technology 11,12 and its differentiation with other enzymatic methods. [86][87][88][89][90] In this review article, we give an overview about the application of sortases in different interdisciplinary research elds. Using sortase A, a protein can be ligated to another biomolecule, a small synthetic molecule, a polymer or a surface.…”

Sortase A-mediated crosslinked short-chain dehydrogenases/reductases as novel biocatalysts with improved thermostability and catalytic efficiency

“…Kim et al . suggested that ligation brought target proteins together to form complementary interfaces, promoting higher catalytic efficiency through substrate channelling, high local substrate and enzyme concentrations and compartmentalisation 37 . Aboye et al .…”

Protein nanoparticles as multifunctional biocatalysts and health assessment sensors

“…Additionally, protein engineering has expanded the biological tools available to post-translationally modify these components. The selection of a right modification strategy is critical to preserving the integrity and the function of the resulting PNPs for the desired applications (Figure 2) [34]. …”