DisProt 데이터베이스로부터 수용성 증진 융합 태그로 활용하기 위한 무정형 단백질 발굴: Disorder score의 중요성

초록

The production of recombinant proteins in Escherichia coli is a fundamental technique across diverse fields of biological research. However, recombinant proteins are often expressed in insoluble forms, and enhancing their solubility remains a major challenge. Fusion protein tags can be used to enhance the solubility of proteins prone to aggregation. Intrinsically disordered protein (IDP)–based solubility-enhancing tags, exemplified by the NEXT tag, have recently emerged as a powerful tool for soluble protein expression. In this study, we sought to discover new IDP tags from the DisProt database, a repository of manually curated IDPs. Unlike the NEXT tag, the candidate tags (λN, Phd, NEP, and Arc) failed to promote soluble expression of the two proteins of interest (βTaCA and PETase), yielding primarily insoluble products. Contrary to expectations, the candidate tags exhibited high secondary-structure content, which appeared to account for their failure to express target proteins solubly. The NEXT tag could be clearly distinguished from the other candidate tags based on disorder prediction and scoring results obtained from IUPRed3 and DISpro. Our findings underscore the need to use disorder-scoring tools to screen for potentially effective solubility-enhancing IDP tags from the DisProt database.

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