title: Revisiting and re-engineering the classical zinc finger peptide: consensus peptide-1 (CP-1).
abstract: Zinc plays key structural and catalytic roles in biology. Structural zinc sites are often referred to as zinc finger (ZF) sites, and the classical ZF contains a Cys2His2 motif that is involved in coordinating Zn(II). An optimized Cys2His2 ZF, named consensus peptide 1 (CP-1), was identified more than 20 years ago using a limited set of sequenced proteins. We have reexamined the CP-1 sequence, using our current, much larger database of sequenced proteins that have been identified from high-throughput sequencing methods, and found the sequence to be largely unchanged. The CCHH ligand set of CP-1 was then altered to a CAHH motif to impart hydrolytic activity. This ligand set mimics the His2Cys ligand set of peptide deformylase (PDF), a hydrolytically active M(II)-centered (M = Zn or Fe) protein. The resultant peptide [CP-1(CAHH)] was evaluated for its ability to coordinate Zn(II) and Co(II) ions, adopt secondary structure, and promote hydrolysis. CP-1(CAHH) was found to coordinate Co(II) and Zn(II) and a pentacoordinate geometry for Co(II)-CP-1(CAHH) was implicated from UV-vis data. This suggests a His2Cys(H2O)2 environment at the metal center. The Zn(II)-bound CP-1(CAHH) was shown to adopt partial secondary structure by 1-D (1)H NMR spectroscopy. Both Zn(II)-CP-1(CAHH) and Co(II)-CP-1(CAHH) show good hydrolytic activity toward the test substrate 4-nitrophenyl acetate, exhibiting faster rates than most active synthetic Zn(II) complexes.
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD 21201, USA. smichel@rx.umaryland.edu.
authors.forename: Angelique N
authors.initials: AN
authors.lastname: Besold
authors.forename: Leland R
authors.initials: LR
authors.lastname: Widger
authors.forename: Frances
authors.initials: F
authors.lastname: Namuswe
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Michalek
authors.forename: Sarah L J
authors.initials: SL
authors.lastname: Michel
authors.forename: David P
authors.initials: DP
authors.lastname: Goldberg
mesh_terms: D000595:Amino Acid Sequence; D017124:Conserved Sequence; D003300:Copper / Q000737:chemistry; D006868:Hydrolysis; D007477:Ions / Q000737:chemistry; D008670:Metals / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D056510:Position-Specific Scoring Matrices; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: C106243:CP-1 peptide; D007477:Ions; D008670:Metals; D009842:Oligopeptides; D010455:Peptides; D003300:Copper; D015032:Zinc