A hemagglutinin from northeast red beans with immunomodulatory activity and anti-proliferative and apoptosis-inducing activities toward tumor cells

Protein Pept Lett. 2013 Oct;20(10):1159-69. doi: 10.2174/0929866511320100011.

Abstract

A 64-kDa hemagglutinin from a Phaseolus vulgaris cultivar, the northeast red bean, was purified by a protocol composed of three chromatographic steps involving affinity chromatography on Affi-gel blue gel, cation exchange chromatography on SP-Sepharose and FPLC-gel filtration on Superdex 75. The purified hemagglutinin appeared as a single 32-kDa band in SDS-PAGE indicating its dimeric nature. The N-terminal amino acid sequence of the hemagglutinin resembled the sequences of lectins and hemagglutinins from a number of Phaseolus species. The hemagglutinin manifested moderate thermostability and pH stability. It retained full activity up to 65 °C and in the pH range 2-12. It did not interact with simple sugars such as glucose, mannose and galactose. The hemagglutinin exerted immunostimulatory effects by upregulating the expression of cytokines like interferon-γ and tumor necrosis factor-α. It also exhibited antiproliferative activity on a number of tumor cells including MCF7 (breast cancer), HepG2 (liver cancer), CNE1 and CNE2 (nasopharyngeal cancer) cells, with stronger activity toward MCF7 and CNE1 cells. The hemagglutinin induced phophatidylserine externalization, mitochondrial depolarization and DNA condensation in MCF7 cells, indicating initiation of apoptosis. However, at high hemagglutinin concentrations, severe damage to the MCF7 cells was detected.

MeSH terms

  • Animals
  • Antineoplastic Agents, Phytogenic / isolation & purification
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apoptosis / drug effects*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Hemagglutinins / isolation & purification
  • Hemagglutinins / pharmacology*
  • Hep G2 Cells
  • Humans
  • Immunologic Factors / isolation & purification
  • Immunologic Factors / pharmacology*
  • Interferon-gamma / genetics
  • MCF-7 Cells
  • Mice
  • Mice, Inbred BALB C
  • Neoplasms / drug therapy
  • Phaseolus / chemistry*
  • Rabbits
  • Tumor Necrosis Factor-alpha / genetics
  • Up-Regulation / drug effects

Substances

  • Antineoplastic Agents, Phytogenic
  • Hemagglutinins
  • Immunologic Factors
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma