The Zinc Antiviral Protein (ZAP) discovered in 2002, has been found to inhibit viral infection by approximately 30 fold in rat cells that express it.  ZAP contains 796 amino acid residues and consists of four CCCH zinc-finger motifs in the N-terminal domain. After ZAP was cloned as a Glutathione S Transferase (GST) fusion protein that produced insoluble protein, the N-terminal domain was re-cloned with maltose binding protein (MBP) or Nus A fusion proteins.  Expression and purification of these fusion proteins was successful, however subsequent cleavage of the solubility tags MBP or Nus A, resulted in precipitation of the ZAP NTD.  ZAP was then re-cloned as a smaller segment fused to MBP or Nus A. The fusion proteins were successfully over expressed, purified and cleaved, producing soluble protein-of-interest.  Buffers are being screened now for optimal solubility and purification of the zinc-fingers of ZAP.

Purification of zinc-fingers 1 & 2 of zinc-antiviral protein (ZAP) via immobilized metal ion affinity chromatography

243rd National Meeting (2012)

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