Selection of a carbohydrate-binding domain with a helix-loop-helix structure

Teruhiko Matsubara, Mie Iida, Takeshi Tsumuraya, Ikuo Fujii, Toshinori Sato

研究成果: Article

17 引用 (Scopus)

抄録

We obtained a novel carbohydrate-binding peptide having a helix-loop-helix scaffold from a random peptide library. The helix-loop-helix peptide library randomized at five amino acid residues was displayed on the major coat protein of a filamentous phage. Affinity selection with a ganglioside, Galβ1-3GalNAcβ1-4(Neu5Acα2-3)Galβ1-4Glcβ1-1′Cer (GM1), gave positive phage clones. Surface plasmon resonance spectroscopy showed that a corresponding 35-mer synthetic peptide had high affinity for GM1 with a dissociation constant of 0.24 μM. This peptide preferentially binds to GM1 rather than asialo GM1 and GM2, suggesting that a terminal galactose and sialic acid are required for the binding as for cholera toxin. Circular dichroism spectroscopic studies indicated that a helical structure is important for the affinity and specificity. Furthermore, alanine scanning at randomized positions showed that arginine and phenylalanine play an especially important role in the recognition of carbohydrates. Such a de novo helix-loop-helix peptide would be available for the design of carbohydrate-binding proteins.

元の言語English
ページ(範囲)6745-6751
ページ数7
ジャーナルBiochemistry
47
発行部数26
DOI
出版物ステータスPublished - 2008 7 1

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Peptide Library
Carbohydrates
Bacteriophages
Peptides
Surface Plasmon Resonance
Gangliosides
Cholera Toxin
Capsid Proteins
N-Acetylneuraminic Acid
Circular Dichroism
Phenylalanine
Galactose
Alanine
Arginine
Spectrum Analysis
Surface plasmon resonance
Clone Cells
Scaffolds
Amino Acids
Spectroscopy

ASJC Scopus subject areas

  • Biochemistry

これを引用

Selection of a carbohydrate-binding domain with a helix-loop-helix structure. / Matsubara, Teruhiko; Iida, Mie; Tsumuraya, Takeshi; Fujii, Ikuo; Sato, Toshinori.

:: Biochemistry, 巻 47, 番号 26, 01.07.2008, p. 6745-6751.

研究成果: Article

Matsubara, Teruhiko ; Iida, Mie ; Tsumuraya, Takeshi ; Fujii, Ikuo ; Sato, Toshinori. / Selection of a carbohydrate-binding domain with a helix-loop-helix structure. :: Biochemistry. 2008 ; 巻 47, 番号 26. pp. 6745-6751.
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