Spectroscopic studies of wild-type and mutant zinc finger peptides: Determinants of domain folding and structure

Journal Article · Mon Jan 01 04:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.1.137· OSTI ID:6810841

Parraga, G; Young, E T; Klevit, R E ^[1]; Horvath, S; Hood, L ^[2]

---

1. Univ. of Washington, Seattle (USA)
2. California Institute of Technology, Pasadena (USA)

The zinc finger model makes both specific structural and specific functional predictions about zinc finger consensus sequences that can be tested with a combination of genetic, molecular biological, and biophysical techniques. The yeast transcription factor ADR1 contains two adjacent zinc finger domains; genetic and deletions in the zinc finger domains resulted in the loss of protein activity. To test the structural and folding predictions of the zinc finger model, peptides encompassing each of the ADR1 fingers were synthesized (ADR1a and ADR1b) as well as a mutant finger peptide (del138) deleted for a single amino acid residue. The folding and metal-binding characteristics of these were assessed by {sup 1}H nuclear magnetic resonance (NMR) and visible spectroscopy. While a single unique conformational species was detected for the two wild-type peptides upon tetrahedral binding of zinc, the deletion peptide did not bind zinc with tetrahedral geometry, nor did it fold into a zinc finger domain. The metal-binding and folding results found with the mutant peptide were similar to those obtained when thiol alkylation or imidazole protonation of the wild-type peptides were performed. These data indicate that ligand spacing and both thiol and imidazole participation in zinc binding are specific and necessary requirements for zinc finger folding, which provides direct support for the initial predictions of the model.

• OSTI ID: 6810841
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 87:1; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ALKYLATION
AMINO ACID SEQUENCE
BARYONS
CHEMICAL REACTIONS
CONFORMATIONAL CHANGES
ELEMENTARY PARTICLES
ELEMENTS
FERMIONS
HADRONS
LIGANDS
MAGNETIC RESONANCE
METALLOPROTEINS
METALS
MOLECULAR STRUCTURE
NMR SPECTRA
NUCLEAR MAGNETIC RESONANCE
NUCLEONS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDES
PROTEIN ENGINEERING
PROTEIN STRUCTURE
PROTEINS
PROTONS
RESONANCE
SPECTRA
THIOLS
TRANSCRIPTION FACTORS
ZINC