Substrate profiling of human transglutaminase 1 using cDNA display and next-generation sequencing

Area of Science:

• Biochemistry
• Molecular Biology
• Dermatology

Background:

• Human transglutaminase 1 (TG1) plays a key role in skin development.
• The specific substrate interactions of TG1 in disease are not well understood.
• Comprehensive exploration of TG1 substrate profile is needed.

Purpose of the Study:

• To investigate the substrate profile of human transglutaminase 1 (TG1).
• To identify specific amino acid sequences recognized and modified by TG1.
• To lay the groundwork for understanding TG1's role in skin disorders.

Main Methods:

• Utilized an in vitro selection system employing cDNA display technology.
• Screened two peptide libraries with varying mutations near the reactive glutamine residue.
• Analyzed results using next-generation sequencing and bioinformatics.

Main Results:

• A detailed TG1 substrate profile was elucidated, highlighting preferred and non-preferred amino acid sequences.
• Identified a highly reactive and specific peptide sequence: AEQHKLPSKWPF.
• Developed a position weight matrix to search human proteins, identifying six known TG1 substrates and candidate substrates.

Conclusions:

• The identified TG1 substrate profile provides insights into its enzymatic activity.
• Findings are expected to aid in future medical diagnoses for skin disorders.
• This research supports the development of novel treatments for skin conditions related to TG1 function.