Advertisement

Apoptosis

, Volume 4, Issue 5, pp 357–362 | Cite as

Mechanism of apoptosis induced by a lysosomotropic agent, L-Leucyl-L-Leucine methyl ester

  • Takayuki Uchimoto
  • Hiroyuki Nohara
  • Rieko Kamehara
  • Michiko Iwamura
  • Naoko Watanabe
  • Yoshiro Kobayashi
Article

Abstract

Lysosomes are fundamental for cell growth, and thus inhibition of the lysosomal function often leads to cell death. L-Leucyl-L-leucine methyl ester (LeuLeuOMe) is a lysosomotropic agent that induces apoptosis of certain immune cells. LeuLeuOMe is taken up through receptor-mediated endocytosis, and then converted into (LeuLeu)n-OMe (n>3) by dipeptidyl peptidase I (DPPI) in lysosomes, which reportedly causes rupture of the lysosomes and DNA fragmentation. In this study we examined how lysosomal damage causes DNA fragmentation in LeuLeuOMe-treated HL-60 cells. When acridine orange was employed to monitor lysosomal membrane integrity, orange or red granular fluorescence was seen in normal cells. In contrast, LeuLeuOMe-treated cells showed orange, yellow or green cellular fluorescence all over the cytoplasm, suggesting that LeuLeuOMe induced a loss of lysosomal membrane integrity. The loss was inhibited by a DPPI inhibitor, GlyPheCHN2 (GFCHN2), but not by a caspase-3 inhibitor, Ac-DEVD-CHO, indicating that a condensation product was responsible for the loss. LeuLeuOMe also induced the activation of caspase-3-like protease and DNA fragmentation, both of which were inhibited by either GFCHN2 or Ac-DEVD-CHO. Therefore, the activation of caspase-3-like protease links the loss of lysosomal membrane integrity to DNA fragmentation during apoptosis induced by LeuLeuOMe.

Acridine orange caspase-3 DNA fragmentation LeuLeuOMe lysosome. 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Neufeld EF, Lim TW, Shapiro LJ. Inherited disorders of lysosomal metabolism. Annu Rev Biochem 1975; 44: 357–376.Google Scholar
  2. 2.
    Nishihara T, Akifusa S, Koseki T, et al. Increase in Bcl-2 level promoted by CD40 ligation correlates with inhibition of B cell apoptosis induced by vacuolar type H+-ATPase inhibitor. Biochem Biophys Res Commun 1995; 212: 255–262.Google Scholar
  3. 3.
    Monney L, Olivier R, Otter I, Jansen B, Poirier GG, Borner C. Role of an acidic compartment in tumor-necrosis-factor-α-induced production of ceramide, activation of caspase-3 and apoptosis. Eur J Biochem 1998; 251: 295–303.Google Scholar
  4. 4.
    Thiele DL, Lipsky PE. The action of leucyl-leucine methyl ester on cytotoxic lymphocytes requires uptake by a novel dipeptide-specific facilitated transfer system and dipeptidyl peptidase I-mediated conversion to membranolytic products. J Exp Med 1990; 172: 183–194.Google Scholar
  5. 5.
    Thiele DL, Lipsky PE. Apoptosis is induced in cells with cytolytic potential by L-Leucyl-L-leucine methyl ester. J Immunol 1992; 148: 3950–3957.Google Scholar
  6. 6.
    Nicholson DW, Thornberry NA. Caspases: killer proteases. TIBS 1997; 22: 299–306.Google Scholar
  7. 7.
    Susin SA, Zamzami N, Castedo M, et al. The central executioner of apoptosis: multiple connections between protease activation and mitochondria in Fas/APO-1/CD95-and ceramide-induced apoptosis. J Exp Med 1997; 186: 25–37.Google Scholar
  8. 8.
    Sakahira H, Enari M, Nagata S. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 1998; 391: 96–99.Google Scholar
  9. 9.
    Odaka M, Furuta T, Kobayashi Y, Iwamura M. Synthesis of caged compounds of L-Leucyl-L-leucine methyl ester, an apoptosis inducer, and their cytotoxic activity. Biochem Biophys Res Commun 1995; 213: 652–656.Google Scholar
  10. 10.
    McGuire MJ, Lipsky PE, Thiele DL. Purification and characterization of dipeptidyl peptidase I from human spleen. Arch Biochem Biophys 1992; 295: 280–288.Google Scholar
  11. 11.
    Nicholson DW, Ali A, Thornberry NA, et al. Identification and inhibition of the ICE/ced-3 protease necessary for mammalian apoptosis. Nature 1995; 376: 37–43.Google Scholar
  12. 12.
    Li W, Yuan XM, Olsson AG, Brunk UT. Uptake of oxidized LDL by macrophages results in partial lysosomal enzyme inactivation and relocation. Arterioscler Thromb Vasc Biol 1998; 18: 177–184.Google Scholar
  13. 13.
    Abrams JM, White K, Fessler LI, Steller H. Programmed cell death during Drosophila embryogenesis Development 1993; 117: 29–43.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Takayuki Uchimoto
    • 1
  • Hiroyuki Nohara
    • 1
  • Rieko Kamehara
    • 1
  • Michiko Iwamura
    • 1
  • Naoko Watanabe
    • 1
  • Yoshiro Kobayashi
    • 1
  1. 1.Department of Biomolecular Science, Faculty of ScienceToho UniversityFunabashi, ChibaJapan

Personalised recommendations