Direct and selective immobilization of proteins by means of an inorganic material-binding peptide: discussion on functionalization in the elongation to material-binding peptide.
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.
Yokoo
Nozomi
N
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan.
Togashi
Takanari
T
Umetsu
Mitsuo
M
Tsumoto
Kouhei
K
Hattori
Takamitsu
T
Nakanishi
Takeshi
T
Ohara
Satoshi
S
Takami
Seiichi
S
Naka
Takashi
T
Abe
Hiroya
H
Kumagai
Izumi
I
Adschiri
Tadafumi
T
D000595:Amino Acid Sequence; D006046:Gold / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*; D011126:Polypropylenes / Q000737:chemistry; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013816:Thermodynamics; D015034:Zinc Oxide / Q000737:chemistry*
D055767:Immobilized Proteins; D010455:Peptides; D011126:Polypropylenes; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D006046:Gold; D015034:Zinc Oxide