A partially purified putative iron P type-ATPase mediates Fe3+-transport into proteoliposome.
We report that two fractions containing proteins from rat hepatocyte nuclei, obtained by nondenaturing gel electrophoresis, were able to bind iron and ATP, and to hydrolyze ATP. Electroelution of these two active fractions followed by SDS-PAGE analysis showed an identical protein pattern, each one containing four proteins in a range of 62-80 kDa. Phosphorylated protein bands were also detected in acid gel and disappeared after treatment with hydroxylamine/acetate or KOH, and upon chasing with cold ATP. A proteoliposome system, made by the incorporation of these partially purified protein fractions into phosphatidylcholine vesicles, carried out Fe(3+)-citrate uptake in a Mg(2+)-ATP-dependent way; Fe(3+) accumulation increased with time reaching a plateau in 30 min. Iron uptake was not supported by AMP-PNP, was partially inhibited by orthovanadate and was not affected by a mix of specific inhibitors of known ATPases. These results support our previous hypothesis that a putative nuclear membrane Fe(3+)-ATPase is involved in nuclear iron homeostasis.
Fessel
Melissa R
MR
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Caixa Postal 26077, CEP 05513-970, São Paulo-SP, Brazil.
Vasconcelos
Eveline G
EG
Gurgueira
Sonia A
SA
Meneghini
Rogério
R
D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005290:Ferric Compounds / Q000378:metabolism*; D022781:Hepatocytes / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000201:enzymology; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D011510:Proteolipids / Q000378:metabolism*; D051381:Rats
D005290:Ferric Compounds; D009687:Nuclear Proteins; D011510:Proteolipids; C020485:proteoliposomes; C025314:ferric citrate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C409468:Fe-ATPase