Engineering of a human kringle domain into agonistic and antagonistic binding proteins functioning in vitro and in vivo

Lee C.-H.; Park K.-J.; Sung E.-S.Kim A.Choi J.-D.Kim J.-S.Kim S.-H.Kwon M.-H.Kim Y.-S.

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Engineering of a human kringle domain into agonistic and antagonistic binding proteins functioning in vitro and in vivo

机译：Engineering of a human kringle domain into agonistic and antagonistic binding proteins functioning in vitro and in vivo

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Here, we report the development of target-specific binding proteins based on the kringle domain (KD) (～80 residues), a ubiquitous modular structural unit occurring across eukaryotic species. By exploiting the highly conserved backbone folding by core residues, but using extensive sequence variations in the seven loop regions of naturally occurring human KDs, we generated a synthetic KD library on the yeast cell surface by randomizing 45 residues in the loops of a human KD template. We isolated KD variants that specifically bind to anticancer target proteins, such as human death receptor 4 (DR4) and/or DR5, and that function as agonists to induce apoptotic cell death in several cancer cell lines in vitro and inhibit tumor progression in mouse models. Combined treatments with KD variants possessing different recognition sites on the same target protein exerted synergisitic tumoricidal activities, compared to treatment with individual variants. In addition to the agonists, we isolated an antagonistic KD variant that binds human tumor necrosis factor-α (TNFα) and efficiently neutralizes TNFα-induced cytotoxicity in vitro and in vivo. The KD scaffold with seven flexible loops protruding from the central core was strongly sequencetolerant to mutations in the loop regions, offering a potential advantage of distinct binding sites for target recognition on the single domain. Our results suggest that the KD scaffold can be used to develop target-specific binding proteins that function as agonists or antagonists toward given target molecules, indicative of their potential use as biotherapeutics.

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《Proceedings of the National Academy of Sciences of the United States of America》 |2010年第21期|9567-9571|共5页
作者
Lee C.-H.; Park K.-J.; Sung E.-S.Kim A.Choi J.-D.Kim J.-S.Kim S.-H.Kwon M.-H.Kim Y.-S.;
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作者单位

Department of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, South Korea;

Institute of Biomedical Science and Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, South Korea;

Department of Chemistry, Chonnam National University, 300, Yongbong-dong, Buk-gu, Gwangju 500-757, South KoreaDepartment of Microbiology, Ajou University School of Medicine, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, South Korea;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
Alternative protein scaffold; Nonantibody scaffold; Protein engineering;

Engineering of a human kringle domain into agonistic and antagonistic binding proteins functioning in vitro and in vivo

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