The effect of C-terminal helix stabilization on specific DNA binding by monomeric GCN4 peptides

Journal of Peptide Science : an Official Publication of the European Peptide Society
Min ZhangJohn W Taylo

Abstract

DNA binding by a 29-residue, monomeric, GCN4 basic region peptide, GCN4br, as well as by peptide br-C, a monomeric basic-region analogue that is helix stabilized at its C-terminal end by a Lys25. Asp29 side-chain lactam-bridged alanine-rich sequence, was studied at 25 C in an aqueous buffer containing 100 mm NaCl. Mixing of both peptides with duplex DNA containing the cAMP-responsive element (CRE) was accompanied by significant helix stabilization in the peptides, whereas mixing of the peptides with duplex DNA containing a scrambled CRE site was not. Peptide NBD-br-C was synthesized as a fluorescent probe to evaluate these peptide-DNA interactions further. Quantitative analysis of the fluorescence quenching of peptide NBD-br-C by CRE half-site DNA indicated the formation of a 1:1 complex with a dissociation constant of 1.41 +/- 0.22 microm. Competitive displacement fluorescence assays of CRE half-site binding gave dissociation constants of 0.65 +/- 0.09 microm for peptide br-C and 3.9 +/- 0.5 microM for GCN4br, which corresponds to a free energy difference of 1.1 kcal/mol that is attributed to the helix stabilization achieved in peptide br-C. This result indicates that helix initiation by the alpha-helical leucine zipper dimeri...Continue Reading

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Citations

Feb 19, 2015·Chemical Communications : Chem Comm·Jesús MosqueraJosé L Mascareñas
Feb 26, 2015·Organic & Biomolecular Chemistry·Abhishek IyerAnnemieke Madder
Oct 8, 2014·Angewandte Chemie·Timothy A HillDavid P Fairlie
Nov 18, 2006·Angewandte Chemie·Juan B BlancoJosé L Mascareñas
May 8, 2010·Chemical Society Reviews·Elena PazosM Eugenio Vázquez
Dec 16, 2003·Chemical Society Reviews·M Eugenio VázquezJ L Mascareñas
Oct 24, 2017·Journal of the American Chemical Society·Soraya Learte-AymamíJosé L Mascareñas

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