Revolutionary Advances: New Methods to Improve Protein Splicing!

Revolutionary Advances: New Methods to Improve Protein Splicing!

The research team around Prof. Dr. Henning Mootz and doctoral student Christoph Humberg from the University of Münster has made significant advances in the field of protein splicing. This study aims to improve the efficiency of laboratory reactions that are critical to producing complex proteins.

Proteins consist of folded peptide chains formed from amino acids and perform a variety of functions in the human body. A central process in protein research is protein splicing, in which inteins are removed from the peptide chain to ensure the correct folding and function of proteins. In their research, the team identified a problem with split inteins that reduces reaction speed and productivity in the laboratory.

Challenges of misfolding

As part of their investigation, the group found that protein misfolding was the cause of the efficiency problems. They developed a method to prevent these misfoldings. Research clearly shows that split inteins can have wide-ranging applications in basic research as well as biotechnology. This is particularly relevant for the synthesis of chimeric proteins, which consist of two parts, one produced in mammalian cells and the other chemically or from bacterial cells.

A specific intein that was examined is the Aes intein. Both fragments of the cleaved intein were produced in bacterial cells, but the productivity was low. A large portion of one of the fragments appeared in the form of an inactive protein aggregate that exhibited specific misfoldings. Bioinformatic analyzes determined which amino acids are responsible for this misfolding and thus led to the development of optimized point mutations in the intein fragment.

Advances through targeted mutations

Through these targeted mutations, the formation of the aggregates could be almost completely suppressed, which significantly increased the productivity of the cleaved intein. These advances could have far-reaching implications for the practical application of protein splicing techniques. According to a report on PMC Over the last 13 years, there has been increasing use of cleaved inteins in biotechnological applications, particularly for the formation of natural peptide bonds.

Despite these advances, many biotechnologists face limitations that hinder the widespread applicability of protein trans-splicing technologies. A deeper understanding of the structure and function of inteins is required to further optimize these technologies. In this context, an investigation describes PubMed that the application of protein splicing as a tool in biochemical research has expanded in many ways over the last 30 years.

The German Research Foundation financially supported the team's work, which underlines the relevance of the research. The insights into improving the productivity of cleaved inteins resulting from this work could not only help improve existing methods, but also open new avenues for protein chemistry and the development of therapeutic approaches.