Protein trans-splicing as a protein ligation tool to study protein structure and function

Area of Science:

• Biochemistry
• Molecular Biology
• Protein Engineering

Background:

• Conventional heterologous protein production faces limitations.
• Split inteins enable protein trans-splicing (PTS), a powerful ligation technique.
• PTS overcomes barriers in producing complex or modified proteins.

Purpose of the Study:

• To provide an overview of split intein engineering and PTS technology.
• To showcase PTS achievements in protein structure-function analyses.
• To discuss factors for successful PTS implementation and future directions.

Main Methods:

• Review of current literature on split intein engineering.
• Analysis of PTS applications in protein modification and reconstitution.
• Discussion of challenges and future prospects for PTS.

Main Results:

• Split intein engineering has advanced PTS capabilities.
• PTS facilitates incorporation of diverse protein modifications.
• PTS enables controllable protein reconstitution, segmental labeling, and cyclization.

Conclusions:

• PTS is a versatile tool for protein engineering and analysis.
• Further development is needed to establish PTS as a universal ligation method.
• PTS holds significant potential for future protein research and biotechnology.