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Direct and Selective Immobilization of Proteins by Means of an Inorganic Material-Binding Peptide: Discussion on Functionalization in the Elongation to Material-Binding Peptide

Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan.
The Journal of Physical Chemistry B (Impact Factor: 3.3). 01/2010; 114(1):480-486. DOI: 10.1021/jp907731b
Source: PubMed

ABSTRACT Using an artificial peptide library, we have identified a peptide with affinity for ZnO materials that could be used to selectively accumulate ZnO particles on polypropylene-gold plates. In this study, we fused recombinant green fluorescent protein (GFP) with this ZnO-binding peptide (ZnOBP) and then selectively immobilized the fused protein on ZnO particles. We determined an appropriate condition for selective immobilization of recombinant GFP, and the ZnO-binding function of ZnOBP-fused GFP was examined by elongating the ZnOBP tag from a single amino acid to the intact sequence. The fusion of ZnOBP with GFP enabled specific adsorption of GFP on ZnO substrates in an appropriate solution, and thermodynamic studies showed a predominantly enthalpy-dependent electrostatic interaction between ZnOBP and the ZnO surface. The ZnOBP's binding affinity for the ZnO surface increased first in terms of material selectivity and then in terms of high affinity as the GFP-fused peptide was elongated from a single amino acid to intact ZnOBP. We concluded that the enthalpy-dependent interaction between ZnOBP and ZnO was influenced by the presence of not only charged amino acids but also their surrounding residues in the ZnOBP sequence.

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    • "This has induced many ongoing efforts aimed to tackle this problem using very different strategies (Moyle et al., 2006; Saenz et al., 2009; Seo et al., 2009; Villa et al., 2011). For antibody or protein binding on solid phases, peptides have the clear disadvantage over larger proteins of their low affinity for the most common solid phases used for macromolecule immobilization, a problem which has also led to specific and often sophisticated approaches to get around it (Li et al., 2011; Palmieri et al., 1995; Yokoo et al., 2010). So, efforts for more intense developments of peptide-based technologies in immunology need to deal with these important hurdles. "
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