Rational design of a calcium-binding protein.
Rational design of a calcium-binding protein.
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DOI：
10.1021/ja034724x
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发表时间：
2003-04
影响因子：
15
通讯作者：
Wei Yang;L. M. Jones;Leanne Isley;Y. Ye;Hsiau-Wei Lee;Anna L. Wilkins;Zhi-Ren Liu;H. Hellinga;R. Malchow;M. Ghazi;Jenny J. Yang
中科院分区：
文献类型：
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作者：
Wei Yang;L. M. Jones;Leanne Isley;Y. Ye;Hsiau-Wei Lee;Anna L. Wilkins;Zhi-Ren Liu;H. Hellinga;R. Malchow;M. Ghazi;Jenny J. Yang
Calcium ions play key roles as structural components in biomineralization and as a second messenger in signaling pathways. We have introduced a de novo designed calcium-binding site into the framework of a non-calcium-binding protein, domain 1 of CD2. The resulting protein selectively binds calcium over magnesium with calcium-binding affinity comparable to that of natural extracellular calcium-binding proteins (K(d) of 50 microM). This experiment is the first successful metalloprotein design that has a high coordination number (seven) metal-binding site constructed into a beta-sheet protein. Our results demonstrate the feasibility of designing a single calcium-binding site into a host protein, taking into account only local properties of a calcium-binding site obtained by a survey of natural calcium-binding proteins and chelators. The resulting site exhibits strong metal selectivity, suggesting that it should now be feasible to understand and manipulate signaling processes by designing novel calcium-modulated proteins with specifically desired functions and to affect their stability.
