title: The putative "nucleation site" in human H-chain ferritin is not required for mineralization of the iron core.
abstract: It is widely believed that the putative nucleation site (Glu61, Glu64, and Glu67) in mammalian H-chain ferritin plays an important role in mineral core formation in this protein. Studies of nucleation site variant A2 (E61A/E64A/E67A) of H-chain ferritin have traditionally shown impaired iron oxidation activity and mineralization. However, recent measurements have suggested that the previously observed impairment may be due to disruption of the ferroxidase site of the protein since Glu61 is a shared ligand of the ferroxidase and nucleation sites of the protein. This study employed a new nucleation site variant A1 (E64A/E67A) which retains the ferroxidase site ligand Glu61. The data (O(2) uptake, iron binding, and conventional and stopped-flow kinetics measurements) show that variant A1 retains a completely functional ferroxidase site and has iron oxidation and mineralization properties similar to those of the wild-type human H-chain protein. Thus, in contrast to previously published literature, this study demonstrates that the putative "nucleation site" does not play an important role in iron uptake or mineralization in H-chain ferritin.
authors.affiliation: Department of Chemistry, University of New Hampshire, Durham, New Hampshire 03824, USA.
authors.forename: Fadi
authors.initials: F
authors.lastname: Bou-Abdallah
authors.forename: Giorgio
authors.initials: G
authors.lastname: Biasiotto
authors.forename: Paolo
authors.initials: P
authors.lastname: Arosio
authors.forename: N Dennis
authors.initials: ND
authors.lastname: Chasteen
mesh_terms: D000409:Alanine / Q000737:chemistry; D002570:Ceruloplasmin / Q000737:chemistry / Q000378:metabolism; D005290:Ferric Compounds / Q000737:chemistry / Q000378:metabolism; D005293:Ferritins / Q000737:chemistry* / Q000378:metabolism; D005296:Ferrous Compounds / Q000737:chemistry / Q000378:metabolism; D018698:Glutamic Acid / Q000737:chemistry; D006801:Humans; D007501:Iron / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008903:Minerals / Q000378:metabolism*; D010101:Oxygen Consumption; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013053:Spectrophotometry
chemical_list: D005290:Ferric Compounds; D005296:Ferrous Compounds; D008903:Minerals; D021122:Protein Subunits; D011994:Recombinant Proteins; D018698:Glutamic Acid; D005293:Ferritins; D007501:Iron; D002570:Ceruloplasmin; D000409:Alanine