TY - JOUR
T1 - Inhibition of brain cation pump enzyme by in vitro lead ion
T2 - Effects of low level [Pb] and modulation by homogenate
AU - Bertoni, John M.
AU - Sprenkle, Pamela M.
N1 - Funding Information:
This research was supported by Grant ES03 184 I from the National Institute of Environmental Health Sciences, by the Gustavus and Louise Pfeiffer Foundation of Santa Monica, California, and by Jefferson Medical College institutional funds.
PY - 1988/3/30
Y1 - 1988/3/30
N2 - 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.
AB - 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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U2 - 10.1016/0041-008X(88)90029-4
DO - 10.1016/0041-008X(88)90029-4
M3 - Article
C2 - 2832972
AN - SCOPUS:0023939786
SN - 0041-008X
VL - 93
SP - 101
EP - 107
JO - Toxicology and Applied Pharmacology
JF - Toxicology and Applied Pharmacology
IS - 1
ER -