Design, Synthesis, and Self‐Assembly Studies of a Suite of Monodisperse, Facially Amphiphilic, Protein–Dendron Conjugates

Abstract

The custom design of protein–dendron amphiphilic macromolecules is at the forefront of macromolecular engineering. Macromolecules with this architecture are very interesting because of their ability to self‐assemble into various biomimetic nanoscopic structures. However, to date, there are no reports on this concept due to technical challenges associated with the chemical synthesis. Towards that end, herein, a new chemical methodology for the modular synthesis of a suite of monodisperse, facially amphiphilic, protein–dendron bioconjugates is reported. Benzyl ether dendrons of different generations (G1–G4) are coupled to monodisperse cetyl ethylene glycol to form macromolecular amphiphilic activity‐based probes (AABPs) with a single protein reactive functionality. Micelle‐assisted protein labeling technology is utilized for site‐specific conjugation of macromolecular AABPs to globular proteins to make monodisperse, facially amphiphilic, protein–dendron bioconjugates. These biohybrid conjugates have the ability to self‐assemble into supramolecular protein nanoassemblies. Self‐assembly is primarily mediated by strong hydrophobic interactions of the benzyl ether dendron domain. The size, surface charge, and oligomeric state of protein nanoassemblies could be systematically tuned by choosing an appropriate dendron or protein of interest. This chemical method discloses a new way to custom‐make monodisperse, facially amphiphilic, protein–dendron bioconjugates.

Programmed and Sequential Disassembly of Multi‐responsive Supramolecular Protein Nanoassemblies: A Detailed Mechanistic Investigation

“…[43] The size and molecular weight of this protein nanoparticle closely match the size of naturally occurring viral capsids. [43] Although extremely powerful, one of the significant limitations of our previous study is that the custom-designed protein assemblies are static, and they failed to exhibit dynamic behavior. This is a severe limitation considering most of the naturally occurring protein complexes exhibit dynamic behavior.…”

“…In a very recent study, we have shown that MAPLab technology can be used for the design of challenging monodisperse well‐defined protein‐dendron bioconjugates [43] . The size and molecular weight of this protein nanoparticle closely match the size of naturally occurring viral capsids [43] . Although extremely powerful, one of the significant limitations of our previous study is that the custom‐designed protein assemblies are static, and they failed to exhibit dynamic behavior.…”

“…We have previously demonstrated that our technology has the following capabilities; i) it is applicable to an entire family of serine proteases, [41] ii) a wide range of hydrophobic molecules (small amphiphilic molecule library, [41] peptides [42] ) can be site‐specifically conjugated to a protein of interest, iii) produces nearly monodisperse protein assemblies which can be studied using standard biophysical and structural biology tools iv) size, oligomeric state, molecular weight and surface charge of protein assemblies can be systematically tuned. In a very recent study, we have shown that MAPLab technology can be used for the design of challenging monodisperse well‐defined protein‐dendron bioconjugates [43] . The size and molecular weight of this protein nanoparticle closely match the size of naturally occurring viral capsids [43] .…”

“…To address some of the limitations of existing methodologies, our group introduced a new chemical method called "micelle-assisted protein labeling (MAPLab) Technology" for the design of semisynthetic proteins. [41][42][43] We have previously demonstrated that our technology has the following capabilities; i) it is applicable to an entire family of serine proteases, [41] ii) a wide range of hydrophobic molecules (small amphiphilic molecule library, [41] peptides [42] ) can be site-specifically conjugated to a protein of interest, iii) produces nearly monodisperse protein assemblies which can be studied using standard biophysical and structural biology tools iv) size, oligomeric state, molecular weight and surface charge of protein assemblies can be systematically tuned. In a very recent study, we have shown that MAPLab technology can be used for the design of challenging monodisperse well-defined proteindendron bioconjugates.…”

Rational Design of Self-assembling Artificial Proteins Utilizing a Micelle-Assisted Protein Labeling Technology (MAPLabTech): Testing the Scope

“…Therefore, it is important to develop a general methodology that can incorporate a wide range of chemical entities onto the self-assembling proteins without compromising salient features of natural proteins such as a single chemical entity and the presence of well-defined functional groups in the 3d-space. Toward that goal, our group invented a new method called 'Micelle-Assisted Protein Labeling Technology (MAPLabTech) [22][23][24][25][26] . During the last several years, we have utilized MAPLabTech for the design of different families of well-defined monodisperse globular SAPs which include protein amphiphiles 22 , protein-dendron conjugates 23 , protein-peptides conjugates 24 , photo-responsive protein amphiphiles 22 , multi-responsive protein-dendron conjugates 25 , and redox-sensitive protein conjugates 26 .…”