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

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

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)6745-6751
Number of pages7
JournalBiochemistry
Volume47
Issue number26
DOIs
Publication statusPublished - 2008 Jul 1

Fingerprint

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

Cite this

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

In: Biochemistry, Vol. 47, No. 26, 01.07.2008, p. 6745-6751.

Research output: Contribution to journalArticle

Matsubara, Teruhiko ; Iida, Mie ; Tsumuraya, Takeshi ; Fujii, Ikuo ; Sato, Toshinori. / Selection of a carbohydrate-binding domain with a helix-loop-helix structure. In: Biochemistry. 2008 ; Vol. 47, No. 26. pp. 6745-6751.
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