RRC ID 35398
著者 Macomber L, Elsey SP, Hausinger RP.
タイトル Fructose-1,6-bisphosphate aldolase (class II) is the primary site of nickel toxicity in Escherichia coli.
ジャーナル Mol Microbiol
Abstract Nickel is toxic to all forms of life, but the mechanisms of cell damage are unknown. Indeed, environmentally relevant nickel levels (8 µM) inhibit wild-type Escherichia coli growth on glucose minimal medium. The same concentration of nickel also inhibits growth on fructose, but not succinate, lactate or glycerol; these results suggest that fructose-1,6-bisphosphate aldolase (FbaA) is a target of nickel toxicity. Cells stressed by 8 µM Ni(II) for 20 min lost 75% of their FbaA activity, demonstrating that FbaA is inactivated during nickel stress. Furthermore, overexpression of fbaA restored growth of an rcnA mutant in glucose minimal medium supplemented with 4 µM Ni(II), thus confirming that FbaA is a primary target of nickel toxicity. This class II aldolase has an active site zinc and a non-catalytic zinc nearby. Purified FbaA lost 80 % of its activity within 2 min when challenged with 8 µM Ni(II). Nickel-challenged FbaA lost 0.8 zinc and gained 0.8 nickel per inactivated monomer. FbaA mutants (D144A and E174A) affecting the non-catalytic zinc were resistant to nickel inhibition. These results define the primary site of nickel toxicity in E. coli as the class II aldolase FbaA through binding to the non-catalytic zinc site.
巻・号 82(5)
ページ 1291-300
公開日 2011-12-1
DOI 10.1111/j.1365-2958.2011.07891.x
PMID 22014167
PMC PMC3225726
MeSH Carbohydrate Metabolism Culture Media / chemistry Enzyme Inhibitors / metabolism Enzyme Inhibitors / toxicity* Escherichia coli / drug effects* Escherichia coli / enzymology* Escherichia coli / metabolism Fructose-Bisphosphate Aldolase / antagonists & inhibitors* Fructose-Bisphosphate Aldolase / isolation & purification Fructose-Bisphosphate Aldolase / metabolism Kinetics Mutant Proteins / antagonists & inhibitors Mutant Proteins / metabolism Nickel / metabolism Nickel / toxicity*
IF 3.418
引用数 42
WOS 分野 MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
原核生物(大腸菌) JW2093-1?(Keio) JW2011-2?(Keio)