Revisiting and re-engineering the classical zinc finger peptide: Consensus peptide-1 (CP-1)

Angelique Besold, Leland R. Widger, Frances Namuswe, Jamie L. Michalek, Sarah L.J. Michel, David P. Goldberg

Research output: Contribution to journalArticle

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 1H 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.

Original languageEnglish (US)
Pages (from-to)1183-1193
Number of pages11
JournalMolecular BioSystems
Volume12
Issue number4
DOIs
StatePublished - Jan 1 2016
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology

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    Besold, A., Widger, L. R., Namuswe, F., Michalek, J. L., Michel, S. L. J., & Goldberg, D. P. (2016). Revisiting and re-engineering the classical zinc finger peptide: Consensus peptide-1 (CP-1). Molecular BioSystems, 12(4), 1183-1193. https://doi.org/10.1039/c5mb00796h