Binding of mammalian and avian ferritins with biotinylated hemin: demonstration of preferential binding of the H subunit to heme.
The binding of ferritin to heme has been well studied using commercial horse spleen apoferritin, which is almost entirely composed of the L subunit, suggesting that mammalian ferritins bind heme. The present study revealed that both mammalian holoferritins (commercial horse spleen ferritin and purified horse spleen, bovine spleen and canine liver ferritins with L/H subunit ratios of 4.0, 1.1, and 2.3, respectively) and their apoferritins bound biotinylated hemin; apoferritins had higher binding activity than holoferritins, except for canine holo- and apoferritins, which showed the same binding. Bovine ferritin H subunit homopolymers expressed by a baculovirus expression system showed heme binding and had higher binding activity to biotinylated hemin than the L subunit homopolymer expressed by the same system. These bindings were inhibited by heme but not by iron-free or Zn-protoporphyrin IX (Zn-PPIX). Purified chicken liver holoferritin was found to be composed of only H subunits and showed the highest binding activity with biotinylated hemin compared with mammalian holoferritins. The binding of chicken liver holoferritin to biotinylated hemin was also inhibited by heme but not by PPIX or Zn-PPIX. These results indicate that mammalian and avian ferritins bind heme and that the H subunit preferentially recognizes heme.
Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Kitasato University.
Miki
M
Nakai
Naomi
N
Murata
Takayuki
T
Yoda
Yasunaga
Y
Yoshikawa
Kiyotaka
K
Watanabe
Koichi
K
Orino
D000818:Animals; D001665:Binding Sites; D019921:Biotinylation; D002645:Chickens / Q000378:metabolism*; D005293:Ferritins / Q000378:metabolism*; D006427:Hemin / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008322:Mammals / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits; D013154:Spleen / Q000378:metabolism
D021122:Protein Subunits; D006427:Hemin; D005293:Ferritins