Folding and Assembly of Vanilloid Receptor Secondary-Structure Peptide with Hexahistidine Linker at Nickel-Nitrilotriacetic Acid Monolayer for Capsaicin Recognition

Koji Nakano, Jun Horiuchi, Shingo Hirata, Makoto Yamanaka, Toshiki Himeno, Ryoichi Ishimatsu

Research output: Contribution to journalArticle

Abstract

Herein, we report the self-assembly of a synthetic vanilloid receptor (VR) peptide that selectively binds capsaicin. We synthesized a 26-mer peptide - YSEILFFVQS-HHHHHH-LAMGWTNMLY (S3HS4) - comprising two chemoreceptor domains of transient receptor potential channel (TRPV1) linked by a hexahistidine sequence. High-speed atomic force microscopy (AFM) imaging in water revealed that the peptide structures alternated rapidly between wedge shape and linear forms. Circular dichroism spectroscopy showed that 65% of the amide units in the peptide chain adopted an α-helix structure, which was ascribed to the chemoreceptor domains. S3HS4 developed well-packed monolayers at the Ni-treated thiolated nitrilotriacetic acid self-assembled monolayers by chelation of the hexahistidine segment, as characterized by infrared spectroscopy and AFM, which exhibited statistically constant specific height. Therefore, S3HS4 was expected to fold spontaneously upon chelation, and the resulting helix-turn-helix conformers developed films while uniformly oriented: the tilt angle was 69° from the surface normal to the substrate. According to microgravimetric analysis using a quartz crystal microbalance (QCM), the adsorption was 84 ± 47 pmol cm -2 (n = 3), which was almost consistent with the saturation adsorption of an α-helix unit. We also used a QCM to investigate the host-guest reactions of S3HS4 and found that the S3HS4-attached QCM-chip-bound capsaicin with an apparent binding constant of (4.2 ± 3.6) × 10 4 M -1 (n = 4), whereas there was no evidence of binding to vanillin or acetophenone. Two controls - a blank chip without S3HS4 and a chip modified with a single helical peptide (LAMGWTNMLY-HHHHHH) - produced no capsaicin response. To the best of our knowledge, S3HS4 is the first example of a synthetic VR mimic peptide. We believe that the present surface-directed structure-based design can be used to exploit the α-helix bundle in hexahistidine-linked bishelical peptides.

Original languageEnglish
Pages (from-to)2047-2054
Number of pages8
JournalLangmuir
Volume35
Issue number6
DOIs
Publication statusPublished - Feb 12 2019

Fingerprint

His-His-His-His-His-His
TRPV Cation Channels
Capsaicin
folding
Peptides
peptides
Monolayers
assembly
Nickel
nickel
helices
acids
Acids
Quartz crystal microbalances
chemoreceptors
quartz crystals
microbalances
chelation
chips
Chelation

All Science Journal Classification (ASJC) codes

  • Materials Science(all)
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Spectroscopy
  • Electrochemistry

Cite this

Folding and Assembly of Vanilloid Receptor Secondary-Structure Peptide with Hexahistidine Linker at Nickel-Nitrilotriacetic Acid Monolayer for Capsaicin Recognition. / Nakano, Koji; Horiuchi, Jun; Hirata, Shingo; Yamanaka, Makoto; Himeno, Toshiki; Ishimatsu, Ryoichi.

In: Langmuir, Vol. 35, No. 6, 12.02.2019, p. 2047-2054.

Research output: Contribution to journalArticle

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abstract = "Herein, we report the self-assembly of a synthetic vanilloid receptor (VR) peptide that selectively binds capsaicin. We synthesized a 26-mer peptide - YSEILFFVQS-HHHHHH-LAMGWTNMLY (S3HS4) - comprising two chemoreceptor domains of transient receptor potential channel (TRPV1) linked by a hexahistidine sequence. High-speed atomic force microscopy (AFM) imaging in water revealed that the peptide structures alternated rapidly between wedge shape and linear forms. Circular dichroism spectroscopy showed that 65{\%} of the amide units in the peptide chain adopted an α-helix structure, which was ascribed to the chemoreceptor domains. S3HS4 developed well-packed monolayers at the Ni-treated thiolated nitrilotriacetic acid self-assembled monolayers by chelation of the hexahistidine segment, as characterized by infrared spectroscopy and AFM, which exhibited statistically constant specific height. Therefore, S3HS4 was expected to fold spontaneously upon chelation, and the resulting helix-turn-helix conformers developed films while uniformly oriented: the tilt angle was 69° from the surface normal to the substrate. According to microgravimetric analysis using a quartz crystal microbalance (QCM), the adsorption was 84 ± 47 pmol cm -2 (n = 3), which was almost consistent with the saturation adsorption of an α-helix unit. We also used a QCM to investigate the host-guest reactions of S3HS4 and found that the S3HS4-attached QCM-chip-bound capsaicin with an apparent binding constant of (4.2 ± 3.6) × 10 4 M -1 (n = 4), whereas there was no evidence of binding to vanillin or acetophenone. Two controls - a blank chip without S3HS4 and a chip modified with a single helical peptide (LAMGWTNMLY-HHHHHH) - produced no capsaicin response. To the best of our knowledge, S3HS4 is the first example of a synthetic VR mimic peptide. We believe that the present surface-directed structure-based design can be used to exploit the α-helix bundle in hexahistidine-linked bishelical peptides.",
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AU - Hirata, Shingo

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AU - Himeno, Toshiki

AU - Ishimatsu, Ryoichi

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