Rational Design of Semi‐Synthetic Protein Complexes with the Defined Oligomeric State

Abstract

Protein oligomers are ubiquitous in nature and play an essential role in various biological processes. Recently, there is an enormous interest to custom‐make synthetic protein oligomers through the bottom‐up approach. In this regard, genetic methods have made tremendous progress in the last decade. In comparison, only few chemical methods currently exist for this purpose. Herein, we report a modular synthetic strategy for the design of facially amphiphilic globular protein‐synthetic peptide conjugates. The self‐assembly of these bioconjugates is driven by strong hydrophobic interaction of the peptide domain. The size, molecular mass and oligomeric state of semi‐synthetic protein complexes strongly depend on size and surface charge of a globular protein as well as the length of the hydrophobic peptide domain. A systematic structure‐property relationship study of these semi‐synthetic proteins should shed more light on the understanding of various non‐covalent forces, which governs the oligomerization processes of naturally occurring proteins.

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

“…Among the chemical technologies used for the design of well-defined monodisperse globular self-assembling artificial proteins, micelle-assisted protein labeling technology (MAPLabTech) has made enormous progress in the last 5 years [22][23][24][25][26] . The MAPLabTech 1.0 was effectively used for the synthesis of SAPs from the proteins belonging to the serine protease family.…”

“…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 .…”

“…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 . Although extremely powerful, the above method was only applicable to proteins that belong to the serine protease family [22][23][24][25][26] . This is a serious limitation and therefore restricts the use of this method for a variety of applications such as vaccine design 6,7 and antibody-drug conjugates.…”

“…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 . Although extremely powerful, the above method was only applicable to proteins that belong to the serine protease family [22][23][24][25][26] .…”

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

“…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 [41–43] . 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 protein‐dendron bioconjugates [43] .…”