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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

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

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

Topics: Research Article
Publisher: Hindawi Publishing Corporation
OAI identifier: oai:pubmedcentral.nih.gov:3092231
Provided by: PubMed Central

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Citations

  1. (1991). a l e s ,P .T .R i c h a r d s o n ,L .M .R o b e r t s ,H .R .W o o d l a n d
  2. (2003). a n dA .P .U .D eA r a ´ ujo,
  3. (1995). A plant lectin derived from Viscum album induces cytokine gene expression and protein production in cultures of human peripheral blood mononuclear cells,”
  4. (2000). Convenient and efficient in vitro folding of disulfidecontaining globular protein from crude bacterial inclusion bodies,”
  5. (2003). Crystal structure at 3 ˚ A of mistletoe lectin I, a dimeric type-II ribosome-inactivating protein, complexed with galactose,”
  6. (2011). Difficulties and perspectives of immunomodulatory therapy with mistletoe lectins and standardized mistletoe extracts in evidence-based medicine,”
  7. (2000). Estimation of protein secondary structure from circular dichroism spectra: comparison of CONTIN, SELCON, and CDSSTR methods with an expanded reference set,”
  8. (1997). h a n ,M .D eS o u s a ,J .A .C h a d d o c k ,L .M .R o b e rt s ,a n dJ .M . Lord,“Restorationoflectinactivitytoanon-glycosylatedricin B chain mutant by the introduction of a novel N-glycosylation site,”
  9. (1990). Increased secretion of tumor necrosis factor α, interleukin 1, and interleukin 6 by human mononuclear cells exposed to β-galactoside-specific lectin from clinically applied mistletoe extract,”
  10. (1997). Isolation and molecular cloning of a novel type 2 ribosome-inactivating protein with an inactive B chain from elderberry (Sambucus nigra) bark,”
  11. (2008). Low-dose mistletoe lectin-I reduces melanoma growth and spread in a scid mouse xenograft model,”
  12. (1997). Nucleation mechanisms in protein folding,”
  13. (1997). Polynucleotide: adenosine glycosidase activity of ribosome-inactivating proteins: effect on DNA, RNA and poly(A),”
  14. (2004). Ribosome-inactivating proteins,”
  15. (1993). Ricin induces the production of tumour necrosis factor-α and interleukin-1β by human peripheral-blood mononuclear cells,”
  16. (1989). T.Hajto,K.Hostanska,andH.J.Gabius,“Modulatorypotency of the β-galactoside-specific lectin from mistletoe extract (Iscador) on the host defense system in vivo in rabbits and patients,”
  17. (1987). The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. The site and the characteristics of the modification in 28 S ribosomal RNA caused by the toxins,”
  18. (1989). u s s a i n ,C .B o w l e r ,L .M .R o b e r t s ,a n dJ
  19. (1998). V a nD a m m e ,W .J .P e u m a n s ,A .B a r r e ,a n dP .R o u g e , “Plant lectins: a composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles,” Critical Reviews in Plant Sciences,