A novel EP-involved pathway for iron release from soya bean seed ferritin.
Iron in phytoferritin from legume seeds is required for seedling germination and early growth. However, the mechanism by which phytoferritin regulates its iron complement to these physiological processes remains unknown. In the present study, protein degradation is found to occur in purified SSF (soya bean seed ferritin) (consisting of H-1 and H-2 subunits) during storage, consistent with previous results that such degradation also occurs during seedling germination. In contrast, no degradation is observed with animal ferritin under identical conditions, suggesting that SSF autodegradation might be due to the EP (extension peptide) on the exterior surface of the protein, a specific domain found only in phytoferritin. Indeed, EP-deleted SSF becomes stable, confirming the above hypothesis. Further support comes from a protease activity assay showing that EP-1 (corresponding to the EP of the H-1 subunit) exhibits significant serine protease-like activity, whereas the activity of EP-2 (corresponding to the EP of the H-2 subunit) is much weaker. Consistent with the observation above, rH-1 (recombinant H-1 ferritin) is prone to degradation, whereas its analogue, rH-2, becomes very stable under identical conditions. This demonstrates that SSF degradation mainly originates from the serine protease-like activity of EP-1. Associated with EP degradation is a considerable increase in the rate of iron release from SSF induced by ascorbate in the amyloplast (pH range, 5.8-6.1). Thus phytoferritin may have facilitated the evolution of the specific domain to control its iron complement in response to cell iron need in the seedling stage.
Fu
Xiaoping
X
China Agricultural University, Beijing, China.
Deng
Jianjun
J
Yang
Haixia
H
Masuda
Taro
T
Goto
Fumiyuki
F
Yoshihara
Toshihiro
T
Zhao
Guanghua
G
D005293:Ferritins / Q000378:metabolism*; D018525:Germination; D007501:Iron / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D055550:Protein Stability; D012639:Seeds / Q000378:metabolism*; D013025:Glycine max / Q000502:physiology*
D010940:Plant Proteins; C030124:phytoferritin; D005293:Ferritins; D007501:Iron