Activation of Helicobacter pylori VacA toxin by alkaline or acid conditions increases its binding to a 250-kDa receptor protein-tyrosine phosphatase β

Kinnosuke Yahiro, Takuro Niidome, Miyuki Kimura, Tomomitsu Hatakeyama, Haruhiko Aoyagi, Hisao Kurazono, Ken Ichi Imagawa, Akihiro Wada, Joel Moss, Toshiya Hirayama

Research output: Contribution to journalArticle

135 Citations (Scopus)

Abstract

Helicobacter pylori, a Gram-negative gastric bacterium, secretes VacA, a cytotoxin that causes vacuolar degeneration of susceptible cells. Velocity sedimentation analysis showed that treatment of VacA at alkaline pH led to disassembly of VacA oligomers, an observation reported previously for acid- treated VacA. Exposure of VacA to acid or alkali increased its binding to AZ- 521 cells, as shown by indirect immunofluorescence and flow cytometry. Moreover, immunoprecipitates with polyclonal antibodies against VacA from AZ- 521 cells previously exposed to acid- or alkali-treated VacA had a 250-kDa glycoprotein containing galactose-β(1-3)-N-acetylgalactosamine and galactose-β(1-4)-N-acetylglucosamine. p250, purified by chromatography on peanut agglutinin affinity and Superose 6 columns, contained N-terminal and internal amino acid sequences of YRQQRKLVEEIGWSYT and LIIQDHILEATQDDY, respectively. These sequences are identical to those of a receptor protein- tyrosine phosphatase (RPTPβ/PTPζ); in agreement, p250 reacted with anti- human RPTPβ monoclonal antibody. Immunopreeipitation with antihuman RPTPβ antibody of solubilized membrane preparations previously incubated with VacA or heat-inactivated VacA demonstrated that RPTPβ bound native, but not denatured, VacA. Acidic and alkaline treatments were associated with activation of VacA and increased binding to the cell surface RPTPβ.

Original languageEnglish
Pages (from-to)36693-36699
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number51
DOIs
Publication statusPublished - Dec 17 1999
Externally publishedYes

Fingerprint

Protein Tyrosine Phosphatases
Helicobacter pylori
Chemical activation
Alkalies
Galactose
Acids
Peanut Agglutinin
Acetylgalactosamine
Acetylglucosamine
Flow cytometry
Antibodies
Cytotoxins
Chromatography
Sedimentation
Oligomers
Glycoproteins
Bacteria
Indirect Fluorescent Antibody Technique
Gram-Negative Bacteria
Monoclonal Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Activation of Helicobacter pylori VacA toxin by alkaline or acid conditions increases its binding to a 250-kDa receptor protein-tyrosine phosphatase β. / Yahiro, Kinnosuke; Niidome, Takuro; Kimura, Miyuki; Hatakeyama, Tomomitsu; Aoyagi, Haruhiko; Kurazono, Hisao; Imagawa, Ken Ichi; Wada, Akihiro; Moss, Joel; Hirayama, Toshiya.

In: Journal of Biological Chemistry, Vol. 274, No. 51, 17.12.1999, p. 36693-36699.

Research output: Contribution to journalArticle

Yahiro, K, Niidome, T, Kimura, M, Hatakeyama, T, Aoyagi, H, Kurazono, H, Imagawa, KI, Wada, A, Moss, J & Hirayama, T 1999, 'Activation of Helicobacter pylori VacA toxin by alkaline or acid conditions increases its binding to a 250-kDa receptor protein-tyrosine phosphatase β', Journal of Biological Chemistry, vol. 274, no. 51, pp. 36693-36699. https://doi.org/10.1074/jbc.274.51.36693
Yahiro, Kinnosuke ; Niidome, Takuro ; Kimura, Miyuki ; Hatakeyama, Tomomitsu ; Aoyagi, Haruhiko ; Kurazono, Hisao ; Imagawa, Ken Ichi ; Wada, Akihiro ; Moss, Joel ; Hirayama, Toshiya. / Activation of Helicobacter pylori VacA toxin by alkaline or acid conditions increases its binding to a 250-kDa receptor protein-tyrosine phosphatase β. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 51. pp. 36693-36699.
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abstract = "Helicobacter pylori, a Gram-negative gastric bacterium, secretes VacA, a cytotoxin that causes vacuolar degeneration of susceptible cells. Velocity sedimentation analysis showed that treatment of VacA at alkaline pH led to disassembly of VacA oligomers, an observation reported previously for acid- treated VacA. Exposure of VacA to acid or alkali increased its binding to AZ- 521 cells, as shown by indirect immunofluorescence and flow cytometry. Moreover, immunoprecipitates with polyclonal antibodies against VacA from AZ- 521 cells previously exposed to acid- or alkali-treated VacA had a 250-kDa glycoprotein containing galactose-β(1-3)-N-acetylgalactosamine and galactose-β(1-4)-N-acetylglucosamine. p250, purified by chromatography on peanut agglutinin affinity and Superose 6 columns, contained N-terminal and internal amino acid sequences of YRQQRKLVEEIGWSYT and LIIQDHILEATQDDY, respectively. These sequences are identical to those of a receptor protein- tyrosine phosphatase (RPTPβ/PTPζ); in agreement, p250 reacted with anti- human RPTPβ monoclonal antibody. Immunopreeipitation with antihuman RPTPβ antibody of solubilized membrane preparations previously incubated with VacA or heat-inactivated VacA demonstrated that RPTPβ bound native, but not denatured, VacA. Acidic and alkaline treatments were associated with activation of VacA and increased binding to the cell surface RPTPβ.",
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AU - Niidome, Takuro

AU - Kimura, Miyuki

AU - Hatakeyama, Tomomitsu

AU - Aoyagi, Haruhiko

AU - Kurazono, Hisao

AU - Imagawa, Ken Ichi

AU - Wada, Akihiro

AU - Moss, Joel

AU - Hirayama, Toshiya

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