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

Self‐assembling artificial proteins (SAPs) have gained enormous interest in recent years due to their applications in different fields. Synthesis of well‐defined monodisperse SAPs is accomplished predominantly through genetic methods. However, the last decade has witnessed the use of a few chemical technologies for this purpose. In particular, micelle‐assisted protein labeling technology (MAPLabTech) has made huge progress in this area. The first generation MAPLabTech focused on site‐specific labeling of the active‐site residue of serine proteases to make SAPs. Further, this methodology was exploited for labeling of N‐terminal residue of a globular protein to make functional SAPs. In this study, we describe the synthesis of novel SAPs by developing a chemical method for site‐specific labeling of a surface‐exposed cysteine residue of globular proteins. In addition, we disclose the synthesis of redox‐sensitive SAPs and their systematic self‐assembly and disassembly studies using size‐exclusion chromatography. Altogether these studies further expand the scope of MAPLabTech in different fields such as vaccine design, targeted drug delivery, diagnostic imaging, biomaterials, and tissue engineering. Advancement of micelle‐assisted protein labeling technology (MAPLabTech 3.0): In this work, we have demonstrated the synthesis of self‐assembling artificial proteins by site‐specific labeling of a single cysteine residue of globular proteins. This methodology should be broadly applicable to a wide range of natural and recombinant proteins containing an engineered cysteine residue.