Preparation of an antibody that recognizes and neutralizes Dictyostelium differentiation-inducing factor-1

Yuzuru Kubohara, Haruhisa Kikuchi, Koji Nakamura, Yusuke Matsuo, Yoshiteru Oshima

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In the development of the cellular slime mold Dictyostelium discoideum, the differentiation-inducing factor-1 (DIF-1; 1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl)hexan-1-one) plays an important role in the regulation of cell differentiation and chemotaxis; however, the cellular signaling systems involving DIF-1 remain to be elucidated. To obtain a probe for DIF-1, we synthesized a DIF derivative (DIF-1-NH2; 6-amino-1-(3,5-dichloro-2,6-dihydroxy-4-methoxyphenyl)hexan-1-one), and prepared an anti-DIF-1 antibody using a DIF-1-NH2-conjugated macromolecule as the immunogen. A 100-fold dilution of the antibody bound to DIF-1-NH2-conjugated resin, and this binding was inhibited by co-addition of 20 μM DIF-1 or DIF-1-NH2. In a monolayer culture of HM44 cells, a DIF-deficient D. discoideum strain, 0.5 nM exogenous DIF-1 induced stalk cell formation in ∼60% of the cells; this induction was dose-dependently inhibited by the antibody (diluted 12.5- or 25-fold). Furthermore, this inhibition by the antibody was recovered by co-addition of 2.5 or10 nM DIF-1. The results indicate that the anti-DIF-1 antibody recognizes DIF-1 and neutralizes its function.

Original languageEnglish
Pages (from-to)364-369
Number of pages6
JournalBiochemical and biophysical research communications
Volume396
Issue number2
DOIs
Publication statusPublished - 2010 May 28

Keywords

  • Cellular slime mold
  • DIF-1
  • HM44
  • Stalk-cell differentiation

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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