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The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling.

Publication ,  Journal Article
Ray, R; de Ridder, GG; Eu, JP; Paton, AW; Paton, JC; Pizzo, SV
Published in: J Biol Chem
September 21, 2012

GRP78, a molecular chaperone with critical endoplasmic reticulum functions, is aberrantly expressed on the surface of cancer cells, including prostate and melanoma. Here it functions as a pro-proliferative and anti-apoptotic signaling receptor via NH(2)-terminal domain ligation. Auto-antibodies to this domain may appear in cancer patient serum where they are a poor prognostic indicator. Conversely, GRP78 COOH-terminal domain ligation is pro-apoptotic and anti-proliferative. There is no method to disrupt cell-surface GRP78 without compromising the total GRP78 pool, making it difficult to study cell-surface GRP78 function. We studied six cell lines representing three cancer types. One cell line per group expresses high levels of cell-surface GRP78, and the other expresses low levels (human hepatoma: Hep3B and HepG2; human prostate cancer: PC3 and 1-LN; murine melanoma: B16F0 and B16F1). We investigated the effect of Escherichia coli subtilase cytoxin catalytic subunit (SubA) on GRP78. We report that SubA specifically cleaves cell-surface GRP78 on HepG2, 1-LN, and B16F1 cells without affecting intracellular GRP78. B16F0 cells (GRP78(low)) have lower amounts of cleaved cell-surface GRP78. SubA has no effect on Hep3B and PC3 cells. The predicted 28-kDa GRP78 COOH-terminal fragment is released into the culture medium by SubA treatment, and COOH-terminal domain signal transduction is abrogated, whereas pro-proliferative signaling mediated through NH(2)-terminal domain ligation is unaffected. These experiments clarify cell-surface GRP78 topology and demonstrate that the COOH-terminal domain is necessary for pro-apoptotic signal transduction occurring upon COOH-terminal antibody ligation. SubA is a powerful tool to specifically probe the functions of cell-surface GRP78.

Duke Scholars

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

September 21, 2012

Volume

287

Issue

39

Start / End Page

32755 / 32769

Location

United States

Related Subject Headings

  • Subtilisins
  • Signal Transduction
  • Receptors, G-Protein-Coupled
  • Proteolysis
  • Prostatic Neoplasms
  • Mice
  • Melanoma
  • Male
  • Humans
  • Hep G2 Cells
 

Citation

APA
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ICMJE
MLA
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Ray, R., de Ridder, G. G., Eu, J. P., Paton, A. W., Paton, J. C., & Pizzo, S. V. (2012). The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling. J Biol Chem, 287(39), 32755–32769. https://doi.org/10.1074/jbc.M112.399808
Ray, Rupa, Gustaaf G. de Ridder, Jerry P. Eu, Adrienne W. Paton, James C. Paton, and Salvatore V. Pizzo. “The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling.J Biol Chem 287, no. 39 (September 21, 2012): 32755–69. https://doi.org/10.1074/jbc.M112.399808.
Ray R, de Ridder GG, Eu JP, Paton AW, Paton JC, Pizzo SV. The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling. J Biol Chem. 2012 Sep 21;287(39):32755–69.
Ray, Rupa, et al. “The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling.J Biol Chem, vol. 287, no. 39, Sept. 2012, pp. 32755–69. Pubmed, doi:10.1074/jbc.M112.399808.
Ray R, de Ridder GG, Eu JP, Paton AW, Paton JC, Pizzo SV. The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling. J Biol Chem. 2012 Sep 21;287(39):32755–32769.

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

September 21, 2012

Volume

287

Issue

39

Start / End Page

32755 / 32769

Location

United States

Related Subject Headings

  • Subtilisins
  • Signal Transduction
  • Receptors, G-Protein-Coupled
  • Proteolysis
  • Prostatic Neoplasms
  • Mice
  • Melanoma
  • Male
  • Humans
  • Hep G2 Cells