Design and Gene Engineering Synthesis of an Extremely Thermostable Protein with Biological Activity

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• 作者：Eiry Kobatake ; Koji Onoda ; Yasuko Yanagida ; and Masuo Aizawa
• 刊名：Biomacromolecules
• 出版年：2000
• 出版时间：September 2000
• 年：2000
• 卷：1
• 期：3
• 页码：382 - 386
• 全文大小：212K
• 年卷期：v.1,no.3(September 2000)
• ISSN：1526-4602

文摘

A novel strategy for designing and synthesizing extremely thermostable biologically active proteins isproposed. The design concept is based on combining a rigid and extremely hydrophobic peptide unit witha biologically active peptide unit. The cell adhesive peptide sequence, Arg-Gly-Asp (RGD), as a functionalpeptide unit was incorporated into the elastin-based rigid polyhexapeptide, whose repeating unit is Ala-Pro-Gly-Val-Gly-Val (APGVGV). The designed fusion gene was expressed in E. coli, and the resultingprotein, designated ER4, was purified with affnity chromatography. The ER4-coated cell culture plate showedsufficient cell adhesive activity through the RGD sequence on the surface of ER4. The thermostability ofER4 was demonstrated by estimating the remaining cell adhesive activity after autoclaving at 120 C for 20min, and it retained over 90% of cell adhesive activity compared with native ER4.