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• W1859396052 abstract "ABSTRACT The cooCTJ gene products are coexpressed with CO-dehydrogenase (CODH) and facilitate in vivo nickel insertion into CODH. A Ni 2+ transport assay was used to monitor uptake and accumulation of 63 Ni 2+ into R. rubrum and to observe the effect of mutations in the cooC , cooT , and cooJ genes on 63 Ni 2+ transport and accumulation. Cells grown either in the presence or absence of CO transported Ni 2+ with a K m of 19 ± 4 μM and a V max of 310 ± 22 pmol of Ni/min/mg of total protein. Insertional mutations disrupting the reading frame of the cooCTJ genes, either individually or all three genes simultaneously, transported Ni 2+ the same as wild-type cells. The nickel specificity for transport was tested by conducting the transport assay in the presence of other divalent metal ions. At a 17-fold excess Mn 2+ , Mg 2+ , Ca 2+ , and Zn 2+ showed no inhibition of 63 Ni 2+ transport but Co 2+ , Cd 2+ , and Cu 2+ inhibited transport 35, 58, and 66%, respectively. Nickel transport was inhibited by cold (50% at 4°C), by protonophores (carbonyl cyanide m -chlorophenylhydrazone, 44%, and 2,4-dinitrophenol, 26%), by sodium azide (25%), and hydroxyl amine (33%). Inhibitors of ATP synthase ( N , N ′-dicyclohexylcarbodiimide and oligomycin) and incubation of cells in the dark stimulated Ni 2+ transport. 63 Ni accumulation after 2 h was four times greater in CO-induced cells than in cells not exposed to CO. The CO-stimulated 63 Ni 2+ accumulation coincided with the appearance of CODH activity in the culture, suggesting that the 63 Ni 2+ was accumulating in CODH. The cooC , cooT , and cooJ genes are required for the increased 63 Ni 2+ accumulation observed upon CO exposure because cells containing mutations disrupting any or all of these genes accumulated 63 Ni 2+ like cells unexposed to CO." @default.
• W1859396052 created "2016-06-24" @default.
• W1859396052 creator A5002590983 @default.
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• W1859396052 date "1999-08-01" @default.
• W1859396052 modified "2023-10-03" @default.
• W1859396052 title "Ni ²⁺ Transport and Accumulation in <i>Rhodospirillum rubrum</i>" @default.
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• W1859396052 doi "https://doi.org/10.1128/jb.181.15.4554-4560.1999" @default.
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