Optimizing Human Epidermal Growth Factor for its Endurance and Specificity Via Directed Evolution: Functional Importance of Leucine at Position 8
Optimizing Human Epidermal Growth Factor for its Endurance and Specificity Via Directed Evolution: Functional Importance of Leucine at Position 8
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通过定向进化优化人表皮生长因子的耐久性和特异性：8位亮氨酸的功能重要性
DOI：
10.1007/s10989-020-10050-6
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发表时间：
2020-02
影响因子：
2.5
通讯作者：
Zhang Qing
中科院分区：
文献类型：
--
作者：
Sun Yujie;Yang Nanyan;Li Fei;Ou Jinxin;Liu Xin;Zhang Qing
表皮生长因子通过结合和激活其受体的胞外区来调节细胞的增殖和迁移。许多工程化的EGF具有增强的激动剂活性，但EGF的半衰期短，对环境的耐受性差，限制了其生理应用。在交错延伸过程中，使用更稳定的猪EGF作为模板，获得了人和猪EGF的两个突变体hL8P和pP8L，并改进了对正常细胞的靶向性。对hEGF和突变体进行了比较研究，以评价它们作为激动剂的活性。突变体对正常细胞和癌细胞的增殖和迁移具有特异性，即pP8L和hL8P均促进正常细胞的增殖和迁移，而延缓癌细胞的增殖和迁移。两种突变体均保持了hEGF的结构热稳定性，pP8L较hEGF具有更强的耐酸性。在HEK293T细胞中的代谢分析表明，hL8P和pP8L突变体的EGFR内化率和生物降解率都较低，但细胞表面的循环率较高。本研究不仅在不降低耐力的情况下提高了EGF激动剂的活性，而且为创伤治疗和肿瘤靶向治疗提供了新的应用前景。
Epidermal growth factor regulates cell proliferation and migration by binding and activating the extracellular region of its receptor. Many engineered EGFs with enhanced agonist activity are produced, yet the short half-life and poor environmental tolerance of EGF limits its physiological application. By using a more stable porcine EGF as the template in the staggered extension process, two mutants of human and porcine EGF, hL8P and pP8L, were obtained with improved targeting towards normal cells. The comparative studies of hEGF and mutants were conducted to evaluate their activities as agonists. The mutants showed specificity for the proliferation and migration of normal and cancer cells, i.e. both pP8L and hL8P promoted the proliferation and migration of normal cells while delayed that of cancer cells. Both mutants preserved the structure thermostability of hEGF and pP8L showed improved acidic tolerance than hEGF. For the metabolism analysis in HEK293T cells, hL8P and pP8L demonstrated the lower rate of both mutants-EGFR internalization and biodegradation, but higher recycling to the cell surface. This study not only enhanced EGF agonist activity without loss of endurance but also provided a new application foreground for trauma therapy and cancer targeting therapy.
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影响因子：
17.1
作者：
Bhirde, Ashwin A.;Patel, Vyomesh;Gavard, Julie;Zhang, Guofeng;Sousa, Alioscka A.;Masedunskas, Andrius;Leapman, Richard D.;Weigert, Roberto;Gutkind, J. Silvio;Rusling, James F.
通讯作者：
Rusling, James F.
影响因子：
16.6
作者：
Kovacs E;Zorn JA;Huang Y;Barros T;Kuriyan J
通讯作者：
Kuriyan J
影响因子：
5.8
作者：
Hector Santana;Yaima González;P. T. Campana;Jesus Noda;Odalys Amarantes;R. Itri;A. Beldarraín;R. Páez
通讯作者：
Hector Santana;Yaima González;P. T. Campana;Jesus Noda;Odalys Amarantes;R. Itri;A. Beldarraín;R. Páez
影响因子：
5.3
作者：
Lu, Chafen;Mi, Li-Zhi;Springer, Timothy A.
通讯作者：
Springer, Timothy A.
影响因子：
14.8
作者：
Greenfield, Norma J.
通讯作者：
Greenfield, Norma J.