Synthesis and Self-Assembly of His-tag Hybrid of Substrate-Binging Short Domain in Transient Receptor Potential Vanilloid Type 1 for Vanillin Sensing Application

• Nakano Koji

  Department of Applied Chemistry, Graduate School of Engineering, Kyushu University

• Hirata Shingo

  Department of Applied Chemistry, Graduate School of Engineering, Kyushu University

• Horiuchi Jun

  Department of Applied Chemistry, Graduate School of Engineering, Kyushu University

• Ishimatsu Ryoichi

  Department of Applied Chemistry, Graduate School of Engineering, Kyushu University

• Imato Toshihiko

  Department of Applied Chemistry, Graduate School of Engineering, Kyushu University

• Onodera Takeshi

  Department of Electrical and Electronic Engineering, Graduate School of Information Science and Electrical Engineering, Kyushu University

• Hayashi Kenshi

  Department of Electrical and Electronic Engineering, Graduate School of Information Science and Electrical Engineering, Kyushu University

抄録

The amino acid sequence of vanillin-binding site of transient receptor potential vanilloid type 1 from rat, Leu⁵⁴⁴–Tyr⁵⁵³, was extracted and hybridized with His-tag. The hexadecamer invariant chain peptide, Leu-Ala-Met-Gly-Trp-Thr-Asn-Met-Leu-Tyr-His-His-His-His-His-His (VBH), was prepared by solid-phase peptide synthesis. Circular dichroic spectral measurements determined the α-helix content to be 17%, which was consistent to that of short peptides. In a combined use of thiol-derivatized nitrilotriacetic acid (s-NTA) monolayers, the His-tag successfully attached the whole peptide on gold substrate surfaces through Ni²⁺-chelation (Γ_VBH = 224 ± 120 pmol cm^-2, n = 8). Moreover, various surface analyses including atomic force microscopy imaging, FT-IR spectroscopy, and quartz-crystal microgravimetry (QCM) revealed self-assembly (SA) of VBH at the S-NTA monolayer surfaces. QCM measurements also showed that vanillin, the major component of natural vanilla flavoring, binds to VBH SAs (K_app = 2.7 × 10³ M^–1). The affinity of host–guest binding remains limited but possesses a certain degree of selectivity; for cases of structural analogs that give a pleasant flavor, acetophenone showed rather weak affinity (K_app = 2.8 × 10² M^–1) whereas 4-heptanone did not bind at all. With these results VBH was concluded to be useful in vanillin sensing as a supramolecular affinity host.

収録刊行物

• Transactions of the Materials Research Society of Japan

  Transactions of the Materials Research Society of Japan 40 (2), 175-178, 2015

  一般社団法人 日本MRS

キーワード

• Transient receptor potential channel
• synthetic peptide
• self-assembly
• biosensor
• vanillin

詳細情報

• CRID: 1390282680490811008
• NII論文ID: 130005089539
• DOI: 10.14723/tmrsj.40.175
• ISSN: 21881650
• ISSN: 13823469
• 本文言語コード: ja
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