High-affinity lead binding proteins in rat kidney cytosol mediate cell-free nuclear translocation of lead.
The PbII binding characteristics of the previously reported (Oskarsson et al., 1982) PbII binding proteins of rat kidney cytosol were investigated further. Saturation and Scatchard analysis of 203Pb binding in whole cytosol and in 40% saturated ammonium sulfate precipitated fractions disclosed a class of relatively high-affinity sites with an apparent Kd of approximately 50 nM and binding capacities of approximately 41 and 9 pmol/mg of protein, respectively. Two 203Pb binding proteins with approximate molecular masses of 63K and 11.5K daltons and a high molecular weight component (greater than 200K) were isolated by Sepharose-6B column chromatography. The time course of association of 203Pb with cytosol and the 63K protein showed maximum binding at 18 hr which was stable up to 25 hr at 4 degrees C. The approximate half-time dissociation rate (T 1/2) of specifically bound 203Pb to the 63K protein was 100 min at 4 degrees C whereas the 11.5K protein showed little dissociation of specifically bound ligand at this temperature. Saturation analysis of the three isolated proteins disclosed low capacity, high-affinity sites with similar apparent Kd values to the cytosol assay. Sucrose density gradient analysis of kidney cytosol showed approximate sedimentation coefficients of 2S, 4.6S and 7S for the 11.5K, 63K and the high molecular weight proteins, respectively. Competitive binding studies with cytosol demonstrated displacement of 203Pb by PbII, CdII and ZnII ions but not CaII ions.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D003600:Cytosol / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007668:Kidney / Q000378:metabolism*; D007854:Lead / Q000378:metabolism*; D008297:Male; D011868:Radioisotopes; D051381:Rats; D011919:Rats, Inbred Strains
D011868:Radioisotopes; D007854:Lead