Expression and Characterization of a Variant of TACI (CRD2-shortTACIFc) in Pichia pastoris

Author(s): Rui Wang, Shiliang Zhou, Xiaomin Peng, Xin-Wen Zhou, Zhi-Qun Xie, Yuxiong Wang, Wei Mo, Min Yu

Journal Name: Protein & Peptide Letters

Volume 19 , Issue 3 , 2012

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TACI is a member of the tumor necrosis factor receptor superfamily and serves as a key regulator of B cell function. The extracellular domain of a typical TNFR contains multiple copies of CRD, which bind in the monomermonomer interfaces of a trimeric ligand. TACI binds to two ligands, APRIL and BAFF, with high affinity and contains two CRD in its extracellular regions, while BCMA and BR3, contain a single or partial CRD for binding the two ligands. However, TACI can be classified as a single CRD receptor because the amino-terminal CRD1 doesnt contribute to ligand binding. To obtain a new variant of TACI possessing higher affinities for binding, we fused a repeat sequence of CRD2 to the N-terminus of the short form of TACI. The new APRIL antagonist peptide, CRD2-shortTACI-Fc, was designed based on the modeling 3-D complex structure of TACI and APRIL. As expected, the purified recombinant CRD2-shortTACI-Fc fusion protein could bind to APRIL in vitro and demonstrated dose-dependent inhibition of APRIL-induced proliferative activity in Raji cells. We found that CRD2-shortTACI-Fc, has a higher affinity for binding to ligands than short-TACI-Fc, which contains a single CRD2.

Keywords: TACI, CRD2, BAFF, APRIL, affinity, autoimmune disorders, Novagen, DNA ligase, homology modeling, Pichia pastoris

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Article Details

Year: 2012
Page: [315 - 325]
Pages: 11
DOI: 10.2174/092986612799363136
Price: $65

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