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; Ha Hyung Kim; Taehoon Chun

doi:10.3109/08923973.2015.1035392

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, Ha Hyung Kim, Taehoon Chun

Research output: Contribution to journal › Article › peer-review

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 E¹²⁵ with D¹²⁵, or E¹²⁵ and C¹²⁷ with D¹²⁵ and F¹²⁷ 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 language	English
Pages (from-to)	221-227
Number of pages	7
Journal	Immunopharmacology and Immunotoxicology
Volume	37
Issue number	3
DOIs	https://doi.org/10.3109/08923973.2015.1035392
Publication status	Published - 2015 Jun 1

Bibliographical note

Funding Information:
The authors declare no conflict of interest. This work was supported by a grant from the Next-Generation BioGreen 21 program (No. PJ011130), Rural Development Administration, Republic of Korea, and by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health &Welfare, Republic of Korea (grant number: HI13C0954).

Publisher Copyright:
© 2015 Informa Healthcare USA, Inc.

Keywords

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

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Toxicology
Pharmacology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.3109/08923973.2015.1035392

Cite this

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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 E125 with D125, or E125 and C127 with D125 and F127 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.",

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author = "Son, {Young Jun} and Jihye Han and Lee, {Jae Yeon} and Kim, {Ha Hyung} and Taehoon Chun",

note = "Funding Information: The authors declare no conflict of interest. This work was supported by a grant from the Next-Generation BioGreen 21 program (No. PJ011130), Rural Development Administration, Republic of Korea, and by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health &Welfare, Republic of Korea (grant number: HI13C0954). Publisher Copyright: {\textcopyright} 2015 Informa Healthcare USA, Inc.",

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AU - Chun, Taehoon

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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