Production of Recombinant Human Transthyretin with Biological Activities toward the Understanding of the Molecular Basis of Familial Amyloidotic Polyneuropathy (FAP)

Hirokazu Furuya, Maria João Mascarenhas Saraiva, Isabel Longo Alves, Mary Ann Gawinowicz, Maria João Mascarenhas Saraiva, Isabel Longo Alves, Pedro P. Costa, Hiroyuki Sasaki, Ikuo Goto, Yoshiyuki Sakaki

研究成果: Contribution to journalArticle査読

74 被引用数 (Scopus)

抄録

Transthyretin (TTR) is a plasma protein interacting with thyroxine T4 and retinol binding protein (RBP). Several variants of TTR with single amino acid substitutions have been identified as the major components of the amyloid fibrils of familial amyloidotic polyneuropathy (FAP), a fetal, autosomal dominant genetic disease. The elucidation of the molecular nature of the variants distinct from that of the wild-type TTR is crucial for understanding the amyloidogenesis in FAP, but our understanding is very poor mainly because of the unavailability of pure variant TTRs. In the present study, we used an Escherichia coli Ompk secretion vector (Ghrayeb et al., 1984) and achieved an effective production of the variant TTRs related to FAP including Met-30, Ile-33, Ala-60, Tyr-77, Met-111, and Ile-122 types. The variant TTRs produced in this system were efficiently secreted to the culture media. The chemical analysis showed that the secreted TTR (Met-30 type) has the same N-terminus as the native one. IEF analyses also indicated that the secreted product is properly processed as assessed by its pI. Furthermore, the secreted TTR was shown to have biological activities, namely, the thyroxin binding activity and the ability to associate with retinol binding protein, indicating that the secreted TTR polypeptide is properly folded. The present work also demonstrated that the processing/secretion of the recombinant TTR molecules in E. coli was strongly affected by single amino acid substitutions.

本文言語英語
ページ(範囲)2415-2421
ページ数7
ジャーナルBiochemistry
30
9
DOI
出版ステータス出版済み - 3 1 1991

All Science Journal Classification (ASJC) codes

  • 生化学

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