Protein domain mapping by λ phage display

The minimal lactose-binding domain of galectin-3

Takanori Moriki, Ichiro Kuwabara, Fu-Tong Liu, Ichi N. Maruyama

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Mapping of protein domains having a distinct function is essential to understanding the protein's structure-function relationship. We used a bacteriophage λ surface expression vector, λfoo, in order to determine the minimal carbohydrate-binding domain of human galectin-3 (Gal-3). Gal-3 cDNA was randomly digested by DNase I and cloned into the phage vector. The library generated was screened by affinity selection using lactose immobilized on agarose beads. DNA sequence analysis of a set of isolated clones defined the minimal folding domain of Gal-3 required for lactose binding, which consisted of 136 amino-acid residues. Using the phage clones isolated, we also determined relative dissociation constants in solution between lactose and the minimal domain expressed on the phage surface. This technique does not require either purified or labeled proteins, and bacteriophage λ surface display may, therefore, be useful for protein domain mapping and in vitro studies of various macromolecular interactions.

Original languageEnglish (US)
Pages (from-to)291-296
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume265
Issue number2
DOIs
StatePublished - Nov 19 1999

Fingerprint

Galectin 3
Bacteriophages
Lactose
Display devices
Proteins
Clone Cells
Deoxyribonuclease I
DNA sequences
DNA Sequence Analysis
Sepharose
Libraries
Membrane Proteins
Complementary DNA
Carbohydrates
Protein Domains
Amino Acids

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Protein domain mapping by λ phage display : The minimal lactose-binding domain of galectin-3. / Moriki, Takanori; Kuwabara, Ichiro; Liu, Fu-Tong; Maruyama, Ichi N.

In: Biochemical and Biophysical Research Communications, Vol. 265, No. 2, 19.11.1999, p. 291-296.

Research output: Contribution to journalArticle

Moriki, Takanori ; Kuwabara, Ichiro ; Liu, Fu-Tong ; Maruyama, Ichi N. / Protein domain mapping by λ phage display : The minimal lactose-binding domain of galectin-3. In: Biochemical and Biophysical Research Communications. 1999 ; Vol. 265, No. 2. pp. 291-296.
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