Open AccessHelicobacter pylori Extracts Exhibit Nicotinamide Adenine Dinucleotide‐derived Adenylation but Not Mono(adenosine 5‐diphosphate‐ribosyOation of DNA Ligase
Author(s) -
Nozaki Tadashige,
Masutani Mitsuko,
Noda Takeshi,
Saito Daizo,
Sugiyama Toshiro,
Takato Tsuyoshi,
Wakabayashi Keiji,
Nakagama Hitoshi,
Sugimura Takashi
Publication year - 1997
Publication title -
japanese journal of cancer research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 0910-5050
DOI - 10.1111/j.1349-7006.1997.tb00308.x
Subject(s) - dna ligase , nad+ kinase , nicotinamide adenine dinucleotide , biochemistry , adenylylation , nicotinamide mononucleotide , helicobacter pylori , biology , microbiology and biotechnology , dna , escherichia coli , enzyme , adenosine diphosphate , chemistry , biosynthesis , genetics , platelet , platelet aggregation , gene , immunology
The issue of toxins produced by Helicobacter pylori ( H. pylori ) urgently requires clarification given that the bacterium causes gastric epithelial cell damage which may lead to precancerous and cancerous changes. During an investigation of the possibility of mono(adenosine 5′‐diphosphate (ADP)‐ribosyl)‐ation by H. pylori products, as observed for other bacterial toxins, we found that radioactivity of [adenylate‐ 32 P]nicotinamide adenine dinucleotide (NAD) is incorporated into an H. pylori protein of 80 kDa after incubation with crude bacterial extract. In contrast, [carbonyl‐ 14 C]NAD did not show any radioactivity incorporation. Unexpectedly, treatment of the modified protein with 0.1 N HC1, but not 0.1 JV NaOH, released the AMP moiety. Such chemical properties are characteristic of bacterial DNA ligase‐AMP complexes. We found that an antibody raised against Escherichia coli DNA ligase [EC 6.5.1.2] immunoprecipitated the modified 80 kDa protein. Our results indicate that incorporation of radioactivity derived from NAD into the 80 kDa protein was due to adenylation, but not mono‐(ADP‐ribosyl) ation, of the DNA ligase of H. pylori.