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The Protein Journal

, Volume 29, Issue 3, pp 174–180 | Cite as

Recombinant Analogs of a Novel Milk Pro-Apoptotic Peptide, Lactaptin, and Their Effect on Cultured Human Cells

  • Dmitry V. Semenov
  • Alexander S. Fomin
  • Elena V. Kuligina
  • Olga A. Koval
  • Vera A. Matveeva
  • Irina N. Babkina
  • Nina V. Tikunova
  • Vladimir A. Richter
Article

Abstract

We recently isolated and characterized a human milk peptide, lactaptin, which induced apoptosis of cultured human MCF-7 cells. Lactaptin was identified as a proteolytic fragment of human kappa-casein. Here, we generated two recombinant analogs of the peptide, RL1 and RL2, containing truncated and complete amino acid sequences of lactaptin, respectively. Analogs were produced in E.coli, purified and assayed for biological activity on cultured human MCF-7 cells. RL1 was shown to induce only a small decrease in cell viability, whereas RL2 lowered the viability of MCF-7 cells by 60%. This reduction in MCF-7 cell viability was associated with apoptosis, which was indicated by phosphatidilserine externalization and caspase-7 activation. The viability of A549 and Hep-2 cells was also reduced by RL2, albeit to a lesser degree than seen with MCF-7 cells; this reduced viability was not accompanied by apoptosis. Non-malignant human mesenchymal stem cells (MSC) were completely resistant to RL2 action.

Keywords

Apoptosis Human kappa-casein Lactaptin MCF-7 

Abbreviations

DFF45

DNA fragmentation factor 45 kDa

FITC

Fluorescein isothiocyanate

MSC

Mesenchymal stem cells

MTT

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PI

Propidium iodide

PS

Phosphatidylserine

TNF-α

Tumor necrosis factor alpha

Notes

Acknowledgments

This work was supported by a grant from the Russian Federal Agency for Science and Innovations, State contract # 02.512.11.2257, #02.522.12.2005 and by an Integration Grant from the Presidium of SB RAS № 18.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Dmitry V. Semenov
    • 1
  • Alexander S. Fomin
    • 1
  • Elena V. Kuligina
    • 1
  • Olga A. Koval
    • 1
  • Vera A. Matveeva
    • 1
  • Irina N. Babkina
    • 1
  • Nina V. Tikunova
    • 1
  • Vladimir A. Richter
    • 1
  1. 1.Institute of Chemical Biology and Fundamental Medicine SB RASNovosibirskRussia

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