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Confining the sodium pump in a phosphoenzyme form: the effect of lead(II) ions.

ABSTRACT: The effect of Pb(2+) ions on the Na(+),K(+)-ATPase was investigated in detail by means of steady-state fluorescence spectroscopy. Experiments were performed by using the electrochromic styryl dye RH421. It is shown that Pb(2+) ions can bind reversibly to the protein and do not affect the Na(+) and K(+) binding affinities in the E(1) and P-E(2) conformations of the enzyme. The pH titrations indicate that lead(II) favors binding of one H(+) to the P-E(2) conformation in the absence of K(+). A model scheme is proposed that accounts for the experimental results obtained for backdoor phosphorylation of the enzyme in the presence of Pb(2+) ions. Taken together, our results clearly indicate that Pb(2+) bound to the enzyme stabilizes an E(2)-type conformation. In particular, under conditions that promote enzyme phosphorylation, Pb(2+) ions are able to confine the Na(+),K(+)-ATPase into a phosphorylated E(2) state.

SUBMITTER: Bartolommei G

PROVIDER: S-EPMC3042568 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature

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Confining the sodium pump in a phosphoenzyme form: the effect of lead(II) ions.

Bartolommei Gianluca G Gramigni Elisa E Tadini-Buoninsegni Francesco F Santini Giacomo G Moncelli Maria Rosa MR

Biophysical journal 20101001 7

The effect of Pb(2+) ions on the Na(+),K(+)-ATPase was investigated in detail by means of steady-state fluorescence spectroscopy. Experiments were performed by using the electrochromic styryl dye RH421. It is shown that Pb(2+) ions can bind reversibly to the protein and do not affect the Na(+) and K(+) binding affinities in the E(1) and P-E(2) conformations of the enzyme. The pH titrations indicate that lead(II) favors binding of one H(+) to the P-E(2) conformation in the absence of K(+). A mode ...[more]

PMID: 20923642

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