Metal-induced folding of a designed metalloprotein.
The metal-induced assembly of a designed peptide-based rubredoxin model is described. The C16C19-GGY peptide has the sequence Ac-K(IEALEGK)(2)(CEACEGK)(IEALEGK)GGY-amide in which the presence of the Cys-X-X-Cys metal binding domain of rubredoxin was used to place cysteine residues at the hydrophobic "a" and "d" positions upon formation of a homodimeric alpha-helical coiled-coil. Circular dichroism spectroscopy shows that the apopeptide exists as a random coil and assembles into a coiled-coil in the presence of Cd(2+). Metal binding is monitored by the appearance of a new LMCT band at 238 nm. UV-Vis titrations and SDS-PAGE experiments are used to show that this designed metalloprotein exists as a metal-bridged coiled-coil dimer.
Kharenko
Olesya A
OA
Department of Chemistry, Center for Photochemical Sciences, Bowling Green State University, 141 Overman, Bowling Green, OH 43403, USA.
Ogawa
Michael Y
MY
D000595:Amino Acid Sequence; D002104:Cadmium; D003198:Computer Simulation; D003545:Cysteine; D019281:Dimerization; D008667:Metalloproteins / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D008670:Metals / Q000737:chemistry* / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017510:Protein Folding
D008667:Metalloproteins; D008670:Metals; D010455:Peptides; D002104:Cadmium; D003545:Cysteine