Abstract
Aim:
To investigate the effect of incorporation of an isoleucine zipper (IZ) motif into CD40 on binding activity of CD40 for the CD40 ligand (CD40L).
Methods:
Prokaryotic expression vectors for 2 soluble CD40 derivatives, shCD40His and shCD40IZ containing an IZ dowain, were constructed and expressed in Escherichia coli. The recombinant proteins were purified to homogeneity after refolding from inclusion bodies. Their molecular weights in solution of shCD40His and shCD40IZ were compared by size-exclusion chromatography, and their binding activity for CD40L on Jurkat T cells was determined by flow cytometry.
Results:
shCD40His and shCD40IZ were generated. Both of them possessed significant binding activity for the cognate ligand CD40L expressed on the cell surface. shCD40IZ had much higher binding activity to its ligand (CD40L) than did shCD40His. Furthermore, size-exclusion chromatography demonstrated that shCD40IZ existed in high molecular mass forms that were most likely to be trimers in solution.
Conclusion:
Incorporation of an IZ motif into CD40 enhances its binding activity for CD40L through trimerization of the CD40 derivative.
Similar content being viewed by others
Article PDF
References
Quezada SA, Jarvinen LZ, Lind EF, Noelle RJ . CD40/CD154 interactions at the interface of tolerance and immunity. Annu Rev Immunol 2004; 22: 307–28.
Xu Y, Song G . The role of CD40-CD154 interaction in cell immunoregulation. J Biomed Sci 2004; 11: 426–38.
Grewal IS, Flavell RA . CD40 and CD154 in cell-mediated immunity. Annu Rev Immunol 1998; 16: 111–35.
Zhang-Hoover J, Sutton A, Stein-Streilein J . CD40/CD40 ligand interactions are critical for elicitation of autoimmune-mediated fibrosis in the lung. J Immunol 2001; 166: 3556–63.
Rossi G, Sarkar J, Scandella D . Long-term induction of immune tolerance after blockade of CD40-CD40L interaction in a mouse model of hemophilia A. Blood 2001; 97: 2750–7.
Homann D, Jahreis A, Wolfe T, Hughes A, Coon B, van Stipdonk MJ, et al. CD40L blockade prevents autoimmune diabetes by induction of bitypic NK/DC regulatory cells. Immunity 2002; 16: 403–15.
Liu H, Mao N, Hou C, Li X, Shen P, Tang PX . Protective effect of human CD40-Ig fusion protein in a murine model of acute graft-versus-host disease. Chin Med J 2001; 114: 685–9.
Wang X, Huang W, Mihara M, Sinha J, Davidson A . Mechanism of action of combined short-term CTLA4Ig and anti-CD40 ligand in murine systemic lupus erythematosus. J Immunol 2002; 168: 2046–53.
Harbury PB, Zhang T, Kim PS, Alber T . A switch between two-, three-, and four-stranded coiled coils in GCN4 leucine zipper mutants. Science 1993; 262: 1401–7.
Sorger PK, Nelson HC . Trimerization of a yeast transcriptional activator via a coiled-coil motif. Cell 1989; 59: 807–13.
Harbury PB, Kim PS, Alber T . Crystal structure of an isoleucine-zipper trimer. Nature 1994; 371: 80–3.
Morris AE, Remmele RL Jr, Klinke R, Macduff BM, Fanslow WC, Armitage RJ . Incorporation of an isoleucine zipper motif enhances the biological activity of soluble CD40L (CD154). J Biol Chem 1999; 274: 418–23.
Shiraishi T, Suzuyama K, Okamoto H, Mineta T, Tabuchi K, Nakayama K, et al. Increased cytotoxicity of soluble Fas ligand by fusing isoleucine zipper motif. Biochem Biophys Res Commun 2004; 322: 197–202.
Tone M, Tone Y, Fairchild PJ, Wykes M, Waldmann H . Regulation of CD40 function by its isoforms generated through alternative splicing. Proc Natl Acad Sci USA 2001; 98: 1751–6.
He XH, Xu LH, Liu Y . Procedure for preparing peptide-major histocompatibility complex tetramers for direct quantification of antigen-specific cytotoxic T lymphocytes. World J Gastroenterol 2005; 11: 4180–7.
Garboczi DN, Hung DT, Wiley DC . HLA-A2-peptide complexes: refolding and crystallization of molecules expressed in Escherichia coli and complexed with single antigenic peptides. Proc Natl Acad Sci USA 1992; 89: 3429–33.
Laemmli UK . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680–5.
Pietravalle F, Lecoanet-Henchoz S, Blasey H, Aubry JP, Elson G, Edgerton MD, et al. Human native soluble CD40L is a biologically active trimer, processed inside microsomes. J Biol Chem 1996; 271: 5965–7.
Mazzei GJ, Edgerton MD, Losberger C, Lecoanet-Henchoz S, Graber P, Durandy A, et al. Recombinant soluble trimeric CD40 ligand is biologically active. J Biol Chem 1995; 270: 7025–8.
Contin C, Pitard V, Delmas Y, Pelletier N, Defrance T, Moreau JF, et al. Potential role of soluble CD40 in the humoral immune response impairment of uraemic patients. Immunology 2003; 110: 131–40.
Holler N, Kataoka T, Bodmer JL, Romero P, Romero J, Deperthes D, et al. Development of improved soluble inhibitors of FasL and CD40L based on oligomerized receptors. J Immunol Methods 2000; 237: 159–73.
Bajorath J, Aruffo A . Construction and analysis of a detailed three-dimensional model of the ligand binding domain of the human B cell receptor CD40. Proteins 1997; 27: 59–70.
Singh J, Garber E, Van Vlijmen H, Karpusas M, Hsu YM, Zheng Z, et al. The role of polar interactions in the molecular recognition of CD40L with its receptor CD40. Protein Sci 1998; 7: 1124–35.
Author information
Authors and Affiliations
Corresponding author
Additional information
Project supported by grants from the National Natural Science Foundation of China (No 30371651 and No 30230350) and National Basic Research Program of China (No G2000057006).
Rights and permissions
About this article
Cite this article
He, Xh., Xu, Lh. & Liu, Y. Enhancement of binding activity of soluble human CD40 to CD40 ligand through incorporation of an isoleucine zipper motif. Acta Pharmacol Sin 27, 333–338 (2006). https://doi.org/10.1111/j.1745-7254.2006.00285.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2006.00285.x
Keywords
This article is cited by
-
An isoleucine-zipper motif enhances costimulation of human soluble trimeric GITR ligand
Cellular & Molecular Immunology (2010)