Plasminogen activator inhibitor-1 fused with erythropoietin (EPO) mimetic peptide (EMP) enhances the EPO activity of EMP

J Pept Res. 2000 Aug;56(2):59-62. doi: 10.1034/j.1399-3011.2000.00716.x.

Abstract

Erythropoietin (EPO) mimetic peptide (EMP) encoding sequence was inserted into the gene of plasminogen activator inhibitor-1 (PAI-1) between Ala348 and Pro349 (P2'-P3'), generating a novel gene, PAI-1/EMP (PMP). This was cloned into pET32a expression vector, fused with TrxA peptide in the vector, and a 63-kDa protein was expressed in inclusion bodies with an expression level >50%. The TrxA/PMP protein was purified by Ni-NTA-agarose metal-ligand affinity chromatography to a purity >90%, showing a single, silver-stained band on SDS-PAGE. Using a reticulocyte counting assay, the EPO activity of PMP was determined to be 5,000 IU/mg, 2,500-fold that of EMP.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chromatography, Affinity
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Erythropoietin / chemistry*
  • Erythropoietin / pharmacology*
  • Escherichia coli / genetics
  • Molecular Mimicry
  • Molecular Sequence Data
  • Peptides / chemistry*
  • Peptides / pharmacology*
  • Plasminogen Activator Inhibitor 1 / chemistry*

Substances

  • DNA Primers
  • Peptides
  • Plasminogen Activator Inhibitor 1
  • Erythropoietin