TY - JOUR
T1 - Cell-cycle inhibition by Helicobacter pylori L-asparaginase
AU - Scotti, Claudia
AU - Sommi, Patrizia
AU - Pasquetto, Maria Valentina
AU - Cappelletti, Donata
AU - Stivala, Simona
AU - Mignosi, Paola
AU - Savio, Monica
AU - Chiarelli, Laurent Roberto
AU - Valentini, Giovanna
AU - Bolanos-Garcia, Victor M.
AU - Merrell, Douglas Scott
AU - Franchini, Silvia
AU - Verona, Maria Luisa
AU - Bolis, Cristina
AU - Solcia, Enrico
AU - Manca, Rachele
AU - Franciotta, Diego
AU - Casasco, Andrea
AU - Filipazzi, Paola
AU - Zardini, Elisabetta
AU - Vannini, Vanio
PY - 2010
Y1 - 2010
N2 - Helicobacter pylori (H. pylori) is a major human pathogen causing chronic gastritis, peptic ulcer, gastric cancer, and mucosa-associated lymphoid tissue lymphoma. One of the mechanisms whereby it induces damage depends on its interference with proliferation of host tissues. We here describe the discovery of a novel bacterial factor able to inhibit the cell-cycle of exposed cells, both of gastric and non-gastric origin. An integrated approach was adopted to isolate and characterise the molecule from the bacterial culture filtrate produced in a protein-free medium: size-exclusion chromatography, non-reducing gel electrophoresis, mass spectrometry, mutant analysis, recombinant protein expression and enzymatic assays. L-asparaginase was identified as the factor responsible for cell-cycle inhibition of fibroblasts and gastric cell lines. Its effect on cell-cycle was confirmed by inhibitors, a knockout strain and the action of recombinant L-asparaginase on cell lines. Interference with cell-cycle in vitro depended on cell genotype and was related to the expression levels of the concurrent enzyme asparagine synthetase. Bacterial subcellular distribution of L-asparaginase was also analysed along with its immunogenicity. H. pylori L-asparaginase is a novel antigen that functions as a cell-cycle inhibitor of fibroblasts and gastric cell lines. We give evidence supporting a role in the pathogenesis of H. pylori-related diseases and discuss its potential diagnostic application.
AB - Helicobacter pylori (H. pylori) is a major human pathogen causing chronic gastritis, peptic ulcer, gastric cancer, and mucosa-associated lymphoid tissue lymphoma. One of the mechanisms whereby it induces damage depends on its interference with proliferation of host tissues. We here describe the discovery of a novel bacterial factor able to inhibit the cell-cycle of exposed cells, both of gastric and non-gastric origin. An integrated approach was adopted to isolate and characterise the molecule from the bacterial culture filtrate produced in a protein-free medium: size-exclusion chromatography, non-reducing gel electrophoresis, mass spectrometry, mutant analysis, recombinant protein expression and enzymatic assays. L-asparaginase was identified as the factor responsible for cell-cycle inhibition of fibroblasts and gastric cell lines. Its effect on cell-cycle was confirmed by inhibitors, a knockout strain and the action of recombinant L-asparaginase on cell lines. Interference with cell-cycle in vitro depended on cell genotype and was related to the expression levels of the concurrent enzyme asparagine synthetase. Bacterial subcellular distribution of L-asparaginase was also analysed along with its immunogenicity. H. pylori L-asparaginase is a novel antigen that functions as a cell-cycle inhibitor of fibroblasts and gastric cell lines. We give evidence supporting a role in the pathogenesis of H. pylori-related diseases and discuss its potential diagnostic application.
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U2 - 10.1371/journal.pone.0013892
DO - 10.1371/journal.pone.0013892
M3 - Article
C2 - 21085483
AN - SCOPUS:78649742504
VL - 5
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 11
M1 - e13892
ER -