Inhibition of brain cation pump enzyme by in vitro lead ion

Effects of low level [Pb] and modulation by homogenate

John M Bertoni, Pamela M. Sprenkle

Research output: Contribution to journalArticle

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Abstract

Rat brain homogenates were preincubated with lead chloride for 20 min at 0°C. Inhibition of K-pnitrophenylphosphatase (K-pNPPase), which measures the dephosphorylation step of Na,K-ATPase, reached steady state within 10 min and was readily reversible. The activity in brain homogenates prepared from 60-day-old rats, expressed (mean ± SE) as a percentage of control (100.0 ± 0.48%), fell to 89.4 ± 0.52% at 0.25 μm [Pb] to 64.2 ± 1.65% at 5 μm [Pb]. However, between 5 and 25 μm [Pb] K-pNPPase activity significantly increased (to 85.4 ± 3.14% at 25 μm) before it again fell above 30 μm. Similar results were obtained with brain homogenates prepared from 10-day-old rats. The multiphasic nature of this dose-response curve did not change with changes in buffer, substrate, anion, test tube order, or test tube composition (glass vs plastic), and appeared to be intrinsic to homogenate-Pb interactions. Microsomal preparations also exhibited multiphasicity, but with a shift in the inflection points. However, there was no multiphasicity with commercial hog brain Na,K-ATPase, suggesting an interaction between a component in the rat brain preparations and lead ion. Addition of boiled rat brain homogenate to commercial Na,K-ATPase resulted in partial protection from lead inhibition but did not produce multiphasicity over the ranges tested. Bovine serum albumin provided less protection. We couclude that there is a complex interaction among K-pNPPase activity, some factor in the rat brain homogenate, and low levels of lead ion.

Original languageEnglish (US)
Pages (from-to)101-107
Number of pages7
JournalToxicology and Applied Pharmacology
Volume93
Issue number1
DOIs
StatePublished - Mar 30 1988

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Cation Transport Proteins
Brain
Rats
Modulation
Ions
Enzymes
Adenosine Triphosphatases
Bovine Serum Albumin
Lead
In Vitro Techniques
Plastics
Glass
Anions
Buffers

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

Cite this

Inhibition of brain cation pump enzyme by in vitro lead ion : Effects of low level [Pb] and modulation by homogenate. / Bertoni, John M; Sprenkle, Pamela M.

In: Toxicology and Applied Pharmacology, Vol. 93, No. 1, 30.03.1988, p. 101-107.

Research output: Contribution to journalArticle

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abstract = "Rat brain homogenates were preincubated with lead chloride for 20 min at 0°C. Inhibition of K-pnitrophenylphosphatase (K-pNPPase), which measures the dephosphorylation step of Na,K-ATPase, reached steady state within 10 min and was readily reversible. The activity in brain homogenates prepared from 60-day-old rats, expressed (mean ± SE) as a percentage of control (100.0 ± 0.48{\%}), fell to 89.4 ± 0.52{\%} at 0.25 μm [Pb] to 64.2 ± 1.65{\%} at 5 μm [Pb]. However, between 5 and 25 μm [Pb] K-pNPPase activity significantly increased (to 85.4 ± 3.14{\%} at 25 μm) before it again fell above 30 μm. Similar results were obtained with brain homogenates prepared from 10-day-old rats. The multiphasic nature of this dose-response curve did not change with changes in buffer, substrate, anion, test tube order, or test tube composition (glass vs plastic), and appeared to be intrinsic to homogenate-Pb interactions. Microsomal preparations also exhibited multiphasicity, but with a shift in the inflection points. However, there was no multiphasicity with commercial hog brain Na,K-ATPase, suggesting an interaction between a component in the rat brain preparations and lead ion. Addition of boiled rat brain homogenate to commercial Na,K-ATPase resulted in partial protection from lead inhibition but did not produce multiphasicity over the ranges tested. Bovine serum albumin provided less protection. We couclude that there is a complex interaction among K-pNPPase activity, some factor in the rat brain homogenate, and low levels of lead ion.",
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