Protein-tyrosine phosphatase alpha, RPTP alpha, is a Helicobacter pylori VacA receptor.
Helicobacter pylori vacuolating cytotoxin, VacA, induces vacuolation, mitochondrial damage, cytochrome c release, and apoptosis of gastric epithelial cells. To detect gastric proteins that serve as VacA receptors, we used VacA co-immunoprecipitation techniques following biotinylation of the cell surface and identified p250, a receptor-like protein-tyrosine phosphatase beta (RPTP beta) as a VacA-binding protein (Yahiro, K., Niidome, T., Kimura, M., Hatakeyama, T., Aoyagi, H., Kurazono, H., Imagawa, K., Wada, A., Moss, J., and Hirayama, T. (1999) J. Biol. Chem. 274, 36693-36699). VacA causes vacuolation of G401 cells, a human kidney tumor cell line, although they do not express RPTP beta. By co-immunoprecipitation with VacA, we identified p140 as a potential receptor in those cells. p140 purified by chromatography on a peanut agglutinin affinity matrix contained internal amino acid sequences of RGEENTDYVNASFIDGYRQK and AEGILDVFQTVK, which are identical to those in RPTP alpha. The peptide mass fingerprinting of p140 by time of flight-MS analysis also supported this identification. Treatment of G401 cells with RPTP alpha-morpholino antisense oligonucleotide before exposure to toxin inhibited vacuolation. These data suggest that RPTP alpha acts as a receptor for VacA in G401 cells. Thus, two receptor tyrosine phosphatases, RPTP alpha and RPTP beta, serve as VacA receptors.
Yahiro
Kinnosuke
K
Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, Nagasaki 8528523, Japan.
Wada
Akihiro
A
Nakayama
Masaaki
M
Kimura
Takahiro
T
Ogushi
Ken-ichi
K
Niidome
Takuro
T
Aoyagi
Haruhiko
H
Yoshino
Ken-ichi
K
Yonezawa
Kazuyoshi
K
Moss
Joel
J
Hirayama
Toshiya
T
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D019921:Biotinylation; D019556:COS Cells; D002846:Chromatography, Affinity; D018922:HL-60 Cells; D016480:Helicobacter pylori / Q000737:chemistry*; D006801:Humans; D007163:Immunosorbent Techniques; D007680:Kidney Neoplasms; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology; D019887:Peanut Agglutinin; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D054630:Receptor-Like Protein Tyrosine Phosphatases, Class 4; D011956:Receptors, Cell Surface; D014162:Transfection; D014407:Tumor Cells, Cultured; D014617:Vacuoles; D009396:Wilms Tumor
D001426:Bacterial Proteins; D016376:Oligonucleotides, Antisense; D019887:Peanut Agglutinin; D010446:Peptide Fragments; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C074689:VacA protein, Helicobacter pylori; C516841:PTPRA protein, human; D017027:Protein Tyrosine Phosphatases; D054630:Receptor-Like Protein Tyrosine Phosphatases, Class 4