Purification and characterization of a low molecular weight zinc binding protein from human placenta.
A low molecular weight, native zinc binding, cytosolic protein (LMZP) has been isolated, purified and characterized from human normal term placenta. Gel filtration of heat treated placental cytosol after sequential acetone precipitation (80% ppt) revealed a major zinc binding protein in the range of low molecular weight. This partially purified zinc binding fraction was further fractionated on DEAE-Sephadex A-25. The zinc was eluted in one of the three peak fractions. Further, the purity of zinc binding protein was confirmed on fast protein liquid chromatography (FPLC). The purified placental LMZP was homogenous on SDS-polyacrylamide gel electrophoresis with a single band. Ultraviolet (UV) spectrum of LMZP showed an absorption maximum at 257 nm which disappeared at pH 2. Molecular weight of LMZP as determined by gel chromatography, SDS-polyacrylamide gel electrophoresis and amino acid analysis was 6 kDa. It was calculated that 1 g atom of zinc was bound to 1 mole of the LMZP. Unlike in classical metallothionein, the amino acid composition of placental LMZP revealed the presence of aromatic amino acids, lower content of cysteine and higher content of histidine, glutamic acid and aspartic acid (10, 9 and 5 residues/mole, respectively).
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Department of Experimental Medicine and Biotechnology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
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D000596:Amino Acids / Q000032:analysis; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel; D004983:Ethanolamines; D005260:Female; D006801:Humans; D008970:Molecular Weight; D010920:Placenta / Q000737:chemistry*; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000737:chemistry / Q000302:isolation & purification*; D015032:Zinc / Q000378:metabolism*
D000596:Amino Acids; D002352:Carrier Proteins; D003911:Dextrans; D004983:Ethanolamines; D011257:Pregnancy Proteins; C017800:zinc-binding protein; C025614:sephadex; D015032:Zinc; C007369:2-diethylaminoethanol