Custom Design of Protein Particles as Multifunctional Biomaterials

Abstract

Assembled protein particles, as emerging biomaterials, have broad applications ranging from vaccines and drug delivery to biocatalysis and particle tracking, but to date these require trial‐and‐error rational design experimentation and/or intensive computational methods to generate. Here, the authors describe an easy‐to‐implement engineering strategy to generate customized protein particles as multifunctional biomaterials. They utilize protein–peptide modules to generate functional nanoparticles whose assembly and size is controlled by the addition of mild stimuli. The protein assembling method is versatile, as exemplified through particle formation with 7 distinct protein modules, using a variety of assembly conditions tailored by the chemistries of 3 peptide partners. They have generated customized protein particles using enzymes, binding and reporter proteins, and their functions and utilities are demonstrated using biocatalysis, sensing, and labelling applications, respectively. Furthermore, co‐assembly with two functional proteins within one particle has been successfully achieved and demonstrated. Physical insights into the kinetics and molecular mechanisms of particle formation are revealed by small angle X‐ray scattering and mass photometry, providing fundamental knowledge to guide design and manufacture these interesting biomaterials in future. Their protein assembling strategy is a reliable method for fabricating a protein particle to deliver new functionalities on‐demand.

Engineering Self-Assembled Endolysin Nanoparticles against Antibiotic-Resistant Bacteria

“…It also enables adequate protein expression levels using common bacterial cultures. Hence, contrary to other proposed approaches, this engineering route is generic, and many proteins can be tagged with this peptide to promote self-organization . P128 endolysin and P 11 4 were connected via a GS linker to facilitate noncovalent peptide interaction and prevent steric hindrance by providing flexibility between protein and peptide motifs .…”

“…Hence, contrary to other proposed approaches, this engineering route is generic, and many proteins can be tagged with this peptide to promote self-organization. 32 P128 endolysin and P 11 4 were connected via a GS linker to facilitate noncovalent peptide interaction and prevent steric hindrance by providing flexibility between protein and peptide motifs. 44 The endolysin−peptide fusion protein (P 11 4−P128) was successfully expressed in recombinant E. coli in high yield, purified and confirmed by SDS-PAGE and liquid chromatography electrospray ionization mass spectrometry (Figure S1, Supporting Information).…”

“…When fused to various functional proteins, these stimuli-responsive peptides confer assembling features to them that promote rapid formation of nanoparticles using diverse assembly conditions such as pH change, salt and temperature. 32 This assembly approach is flexible, straightforward and scalable and does not require extensive protein structural details to promote nanoparticle formation, making these assembly approaches and their related protein-based nanomaterials promising. In this work, endolysin was engineered with a self-assembly peptide to form nanoparticles with inherent bacterial killing capability (Figure 1A).…”

“…In an attempt to increase the biological performance of endolysins, we hypothesized that stimuli-responsive peptides could be utilized to design and self-assemble endolysins into protein-based antibacterial nanoparticles with promising antibacterial properties, which are not owned by the native endolysin building blocks. When fused to various functional proteins, these stimuli-responsive peptides confer assembling features to them that promote rapid formation of nanoparticles using diverse assembly conditions such as pH change, salt and temperature . This assembly approach is flexible, straightforward and scalable and does not require extensive protein structural details to promote nanoparticle formation, making these assembly approaches and their related protein-based nanomaterials promising.…”

Plant Protein‐Peptide Supramolecular Polymers with Reliable Tissue Adhesion for Surgical Sealing

“…[ 33 ] In particular, the catalytic or binding capability of the parent proteins can be well retained within the assembled particles. [ 33 ] As an alternative way, non‐covalent assembly between recombinant elastin‐like protein and amphiphilic peptides has been explored to generate macroscopic membrane and tubular scaffold for guiding cell growth. [ 34 ] Further combing the bottom‐up assembly with 3D‐printing technique, bioactive protein‐peptide scaffolds with higher‐ordered structures at macroscopic level have been constructed for tissue engineering [ 35 ] and multicellular 3D model for ovarian cancer.…”