Purification, electrophoretic behavior, and kinetics of iron release of liver ferritin of Dasyatis akajei.
From the liver of fish Dasyatis akajei, ferritin has been isolated by thermal denaturation and ammonium sulfate fractionation and then further purified by anion exchange chromatography and gel exclusion chromatography. The molecular weight of the liver ferritin of D. akajei (DALF) was measured to be 400 kDa by PAGE. Moreover, SDS-PAGE experimentation indicates that protein shell of DALF consists of the H and L subunits with molecular weight of 18 and 13 kDa, respectively. Using isoelectric focusing with pH ranging from 5.0 to 6.0, the ferritin purified by the PAGE exhibited three bands with different pI values in the gel slab. Diameters of the protein shell and iron core were also investigated by transmission electron microscope and determined to be 10-12 nm and 5-8 nm, respectively. A kinetic study of DALF reveals that the rate of self-regulation of the protein shell rather than the complex surface of the iron core plays an important role in forming a process for iron release with mixed orders.
Kong
Bo
B
Department of Biology, The Center for Analysis and Testing, The Key Laboratory of MOE for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen 361005, China.
Huang
He-Qing
HQ
Lin
Qing-Mei
QM
Kim
Won-Suk
WS
Cai
Zongwei
Z
Cao
Ting-Ming
TM
Miao
Hai
H
Luo
Da-Min
DM
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005290:Ferric Compounds / Q000737:chemistry*; D005293:Ferritins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D005399:Fishes / Q000502:physiology*; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007700:Kinetics; D008099:Liver / Q000737:chemistry*; D008854:Microscopy, Electron; D008970:Molecular Weight; D010710:Phosphates / Q000737:chemistry
D005290:Ferric Compounds; D010710:Phosphates; D005293:Ferritins