Nature Structural Biology
2, 368 - 373 (1995)
doi:10.1038/nsb0595-368
Novel metal-binding proteins by designMichael Klemba1, Kevin H. Gardner1, Stephen Marino1, Neil D. Clarke2
& Lynne Regan1
1Department of Molecular Biophysics & Biochemistry Yale University 266 Whitney Avenue New Haven Connecticut 06520, USA
2Department of Biophysics, Johns Hopkins University, Baltimore, Maryland 21205, USA We describe the successful design of a tetrahedral His3Cys Zn(II)-binding site in a small protein of known structure: the B1 domain of Streptococcal protein G. The B1 variants containing the novel metal-binding site were characterized using a combination of optical absorption, circular dichroism and NMR spectroscopies. The results indicate that the designed proteins bind Zn(II) with high affinity and tetrahedral coordination geometry, and that the overall secondary and tertiary structure of the B1 domain is maintained.
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