title: Biochemical evidence for lead and mercury induced transbilayer movement of phospholipids mediated by human phospholipid scramblase 1.
abstract: Human phospholipid scramblase 1(hPLSCR1) is a transmembrane protein involved in bidirectional scrambling of plasma membrane phospholipids during cell activation, blood coagulation, and apoptosis in response to elevated intracellular Ca(2+) levels. Pb(2+) and Hg(2+) are known to cause procoagulant activation via phosphatidylserine exposure to the external surface in erythrocytes, resulting in blood coagulation. To explore its role in lead and mercury poisoning, hPLSCR1 was overexpressed in Escherichia coli BL21 (DE3) and purified using affinity chromatography. The biochemical assay showed rapid scrambling of phospholipids in the presence of Hg(2+) and Pb(2+). The binding constant (Ka) was calculated and found to be 250 nM(-1) and 170 nM(-1) for Hg(2+) and Pb(2+), respectively. The intrinsic tryptophan fluorescence and far ultraviolet circular dichroism studies revealed that Hg(2+) and Pb(2+) bind to hPLSCR1 and induce conformational changes. hPLSCR1 treated with protein modifying reagent N-ethylmaleimide before functional reconstitution showed 40% and 24% inhibition in the presence of Hg(2+) and Pb(2+), respectively. This is the first biochemical evidence to prove the above hypothesis that hPLSCR1 is activated in heavy metal poisoning, which leads to bidirectional transbilayer movement of phospholipids.
authors: Applied and Industrial Microbiology Laboratory, Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600 036, India.
authors: Ashok Kumar
authors: AK
authors: Shettihalli
authors: Sathyanarayana N
authors: SN
authors: Gummadi
mesh_terms: D006801:Humans; D007854:Lead / Q000494:pharmacology*; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008628:Mercury / Q000494:pharmacology*; D049294:Phospholipid Transfer Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008051:Lipid Bilayers; C493422:PLSCR1 protein, human; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; D007854:Lead; D008628:Mercury