Production of active nonglycosylated recombinant B-chain of type-2 ribosome-inactivating protein from viscum articulatum and its biological effects on peripheral blood mononuclear cells

Ching Hsiang Hsu, Tzu Li Lu, Jing Yuan Chuang, Jai Sing Yang, Shau Ting Chiu, Nai-Wan Hsiao, Mei Chen Wu, Shih Hsiung Wu

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Type-2 ribosome-inactivating proteins, composed of a toxic A-chain and lectin-like B-chain, display various biological functions, including cytotoxicity and immunomodulation. We here cloned the lectin-like B-chain encoding fragment of a newly identified type-2 RIP gene, articulatin gene, from Viscum articulatum, into a bacterial expression vector to obtain nonglycosylated recombinant protein expressed in inclusion bodies. After purification and protein refolding, soluble refolded recombinant articulatin B-chain (rATB) showed lectin activity specific toward galactoside moiety and was stably maintained while stored in low ionic strength solution. Despite lacking glycosylation, rATB actively bound leukocytes with preferential binding to monocytes and in vitro stimulated PBMCs to release cytokines without obvious cytotoxicity. These results implicated such a B-chain fragment as a potential immunomodulator.

Original languageEnglish
Article number283747
JournalEvidence-based Complementary and Alternative Medicine
Volume2011
DOIs
Publication statusPublished - 2011 Jun 22

Fingerprint

Type 2 Ribosome Inactivating Proteins
Viscum
Blood Cells
Protein Refolding
Galactosides
Immunomodulation
Poisons
Inclusion Bodies
Immunologic Factors
Glycosylation
Recombinant Proteins
Lectins
Osmolar Concentration
Genes
Monocytes
Leukocytes
Cytokines
Phytolacca americana lectin B

All Science Journal Classification (ASJC) codes

  • Complementary and alternative medicine

Cite this

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title = "Production of active nonglycosylated recombinant B-chain of type-2 ribosome-inactivating protein from viscum articulatum and its biological effects on peripheral blood mononuclear cells",
abstract = "Type-2 ribosome-inactivating proteins, composed of a toxic A-chain and lectin-like B-chain, display various biological functions, including cytotoxicity and immunomodulation. We here cloned the lectin-like B-chain encoding fragment of a newly identified type-2 RIP gene, articulatin gene, from Viscum articulatum, into a bacterial expression vector to obtain nonglycosylated recombinant protein expressed in inclusion bodies. After purification and protein refolding, soluble refolded recombinant articulatin B-chain (rATB) showed lectin activity specific toward galactoside moiety and was stably maintained while stored in low ionic strength solution. Despite lacking glycosylation, rATB actively bound leukocytes with preferential binding to monocytes and in vitro stimulated PBMCs to release cytokines without obvious cytotoxicity. These results implicated such a B-chain fragment as a potential immunomodulator.",
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Production of active nonglycosylated recombinant B-chain of type-2 ribosome-inactivating protein from viscum articulatum and its biological effects on peripheral blood mononuclear cells. / Hsu, Ching Hsiang; Lu, Tzu Li; Chuang, Jing Yuan; Yang, Jai Sing; Chiu, Shau Ting; Hsiao, Nai-Wan; Wu, Mei Chen; Wu, Shih Hsiung.

In: Evidence-based Complementary and Alternative Medicine, Vol. 2011, 283747, 22.06.2011.

Research output: Contribution to journalArticle

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AU - Hsu, Ching Hsiang

AU - Lu, Tzu Li

AU - Chuang, Jing Yuan

AU - Yang, Jai Sing

AU - Chiu, Shau Ting

AU - Hsiao, Nai-Wan

AU - Wu, Mei Chen

AU - Wu, Shih Hsiung

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