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Recombinant Production and Characterization of SAC, the Core Domain of Par-4, by SUMO Fusion System

Jian Zhang1, Aiyou Sun2, Yuguo Dong1

  • 1State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.

Applied Biochemistry and Biotechnology
|October 4, 2017
PubMed

Insights

Prostate apoptosis response-4 (Par-4) core domain, SAC, effectively inhibits ovarian cancer cell growth and induces apoptosis by binding to GRP78. This study presents an efficient method for producing bioactive SAC for potential cancer therapy.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • Prostate apoptosis response-4 (Par-4) is an anticancer protein targeting cancer cell proliferation and apoptosis.
  • The core domain of Par-4 (SAC) demonstrates tumor growth inhibition without harming normal tissues.
  • The therapeutic potential of SAC in ovarian cancer remains unexplored.

Purpose of the Study:

  • To develop an effective method for producing native SAC in prokaryotic cells.
  • To evaluate the anticancer effects of recombinant SAC on ovarian cancer cells.
  • To elucidate the mechanism of SAC-induced apoptosis in ovarian cancer.

Main Methods:

  • A small ubiquitin-related modifier (SUMO) fusion system was employed for recombinant SAC production in E. coli.
  • Ni-NTA affinity chromatography was used for purification, followed by SUMO protease cleavage.
  • Ovarian cancer cell proliferation and apoptosis assays were performed, along with immunofluorescence and binding studies.

Main Results:

  • A simplified, one-step purification process yielded highly pure (95%) recombinant SAC with a yield of 25 mg/L.
  • Recombinant SAC significantly inhibited proliferation and induced apoptosis in SKOV-3 ovarian cancer cells.
  • SAC specifically binds to the cell surface receptor GRP78 to trigger apoptosis in SKOV-3 cells.

Conclusions:

  • An effective strategy for expressing bioactive SAC in prokaryotic systems was established.
  • Recombinant SAC exhibits potent anticancer activity against ovarian cancer cells.
  • The findings support the potential application of SAC in ovarian cancer therapy.
Keywords:
GRP78One-step purificationOvarian cancerProstate apoptosis response-4 (Par-4)SACSmall ubiquitin-related modifier (SUMO)

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