Improved efficacy of soluble human receptor activator of nuclear factor kappa B (RANK) fusion protein by site-directed mutagenesis

Young Jun Son, Jihye Han, Jae Yeon Lee, HaHyung Kim, Taehoon Chun

Research output: Contribution to journalArticle

Abstract

Soluble human receptor activator of nuclear factor kappa B fusion immunoglobulin (hRANK-Ig) has been considered as one of the therapeutic agents to treat osteoporosis or diseases associated with bone destruction by blocking the interaction between RANK and the receptor activator of nuclear factor kappa B ligand (RANKL). However, no scientific record showing critical amino acid residues within the structural interface between the human RANKL and RANK complex is yet available. In this study, we produced several mutants of hRANK-Ig by replacement of amino acid residue(s) and tested whether the mutants had increased binding affinity to human RANKL. Based on the results from flow cytometry and surface plasmon resonance analyses, the replacement of E<sup>125</sup> with D<sup>125</sup>, or E<sup>125</sup> and C<sup>127</sup> with D<sup>125</sup> and F<sup>127</sup> within loop 3 of cysteine-rich domain 3 of hRANK-Ig increases binding affinity to human RANKL over the wild-type hRANK-Ig. This result may provide the first example of improvement in the efficacy of hRANK-Ig by protein engineering and may give additional information to understand a more defined structural interface between hRANK and RANKL.

Original languageEnglish
Pages (from-to)221-227
Number of pages7
JournalImmunopharmacology and Immunotoxicology
Volume37
Issue number3
DOIs
Publication statusPublished - 2015 Jun 1

Fingerprint

Receptor Activator of Nuclear Factor-kappa B
Mutagenesis
RANK Ligand
Site-Directed Mutagenesis
Fusion reactions
Immunoglobulins
Proteins
UCON 50-HB-5100
Amino Acids
Flow cytometry
Protein Engineering
Surface plasmon resonance
kappa Opioid Receptor
Surface Plasmon Resonance
Cysteine
Bone
Osteoporosis
Flow Cytometry
Bone and Bones

Keywords

  • Immunoglobulin fusion protein
  • Mutagenesis
  • Osteoclast
  • Osteoporosis
  • Receptor activator of nuclear factor kappa B

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Pharmacology
  • Toxicology

Cite this

Improved efficacy of soluble human receptor activator of nuclear factor kappa B (RANK) fusion protein by site-directed mutagenesis. / Son, Young Jun; Han, Jihye; Lee, Jae Yeon; Kim, HaHyung; Chun, Taehoon.

In: Immunopharmacology and Immunotoxicology, Vol. 37, No. 3, 01.06.2015, p. 221-227.

Research output: Contribution to journalArticle

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