Contribution of individual zinc ligands to metal binding and peptide folding of zinc finger peptides.
Little is known about the contribution of individual zinc-ligating amino acid residues for coupling between zinc binding and protein folding in zinc finger domains. To understand such roles of each zinc ligand, four zinc finger mutant peptides corresponding to the second zinc finger domain of Sp1 were synthesized. In the mutant peptides, glycine was substituted for one of four zinc ligands. Their metal binding and folding properties were spectroscopically characterized and compared to those of the native zinc finger peptide. In particular, the electronic charge-transfer and d-d bands of the Co(II)-substituted peptide complexes were used to examine the metal coordination number and geometry. Fluorescence emission studies revealed that the mutant peptides are capable of binding zinc despite removing one ligand. Circular dichroism results clearly showed the induction of an alpha-helix by zinc binding. In addition, the structures of certain mutant zinc finger peptides were simulated by molecular dynamics calculation. The information indicates that His23 and the hydrophobic core formed between the alpha-helix and the beta-sheet play an essential role in alpha-helix induction. This report demonstrates that each ligand does not contribute equally to alpha-helix formation and coordination geometry in the zinc finger peptide.
Nomura
Akiko
A
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
Sugiura
Yukio
Y
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D003035:Cobalt / Q000737:chemistry; D005998:Glycine / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006801:Humans; D008024:Ligands; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D017510:Protein Folding; D016329:Sp1 Transcription Factor / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers
D008024:Ligands; D010455:Peptides; D016329:Sp1 Transcription Factor; D003035:Cobalt; D006639:Histidine; D015032:Zinc; D005998:Glycine