High expression and purification of amino-terminal fragment of human amyloid precursor protein in Pichia pastoris and partial analysis of its properties

Biomed Res Int. 2013:2013:836429. doi: 10.1155/2013/836429. Epub 2013 Nov 7.

Abstract

The cleaved amino-terminal fragment of human amyloid precursor protein (N-APP) binds death receptor 6 (DR6) and triggers a caspase-dependent self-destruction process, which was suggested to contribute to Alzheimer's disease. To investigate the N-APP-DR6-induced degeneration pathway at the molecular level, obtaining abundant and purified N-APP is fundamental and critical. The recombinant N-APP has been produced in mammalian expression system. However, the cost and yield disadvantages of mammalian expression system make it less ideal for protein mass production. Here, we successfully expressed and purified recombinant N-terminal 18-285 amino acid residues of human amyloid precursor protein from the methylotrophic yeast Pichia pastoris with a high yield of 50 mg/L. Flow cytometry indicated the purified N-APP-induced obvious apoptosis of human neuroblastoma SHEP cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid beta-Protein Precursor* / biosynthesis
  • Amyloid beta-Protein Precursor* / chemistry
  • Amyloid beta-Protein Precursor* / genetics
  • Amyloid beta-Protein Precursor* / isolation & purification
  • Amyloid beta-Protein Precursor* / pharmacology
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Gene Expression*
  • Humans
  • Pichia*
  • Protein Structure, Tertiary
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification

Substances

  • APP protein, human
  • Amyloid beta-Protein Precursor
  • Recombinant Proteins