title: Novel metal-binding proteins by design.
abstract: 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.
authors.affiliation: Department of Molecular Biophysics & Biochemistry Yale University, New Haven, Connecticut 06520, USA.
authors.forename: M
authors.initials: M
authors.lastname: Klemba
authors.forename: K H
authors.initials: KH
authors.lastname: Gardner
authors.forename: S
authors.initials: S
authors.lastname: Marino
authors.forename: N D
authors.initials: ND
authors.lastname: Clarke
authors.forename: L
authors.initials: L
authors.lastname: Regan
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism; D002942:Circular Dichroism; D003035:Cobalt / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008667:Metalloproteins / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D013816:Thermodynamics; D015032:Zinc / Q000378:metabolism
chemical_list: D007074:Immunoglobulin G; D008667:Metalloproteins; D002104:Cadmium; D003035:Cobalt; D006639:Histidine; D015032:Zinc; D003545:Cysteine