Rational design of a calcium-binding protein.

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Yang, W; Jones, LM; Isley, L; Ye, Y; Lee, H-W; Wilkins, A; Liu, Z-R; Hellinga, HW; Malchow, R; Ghazi, M; Yang, JJ

Published Date

  • May 21, 2003

Published In

Volume / Issue

  • 125 / 20

Start / End Page

  • 6165 - 6171

PubMed ID

  • 12785848

Pubmed Central ID

  • 12785848

International Standard Serial Number (ISSN)

  • 0002-7863

Digital Object Identifier (DOI)

  • 10.1021/ja034724x

Language

  • eng

Conference Location

  • United States