title: Ferritin protein nanocage ion channels: gating by N-terminal extensions.
abstract: Ferritin protein nanocages, self-assembled from four-α-helix bundle subunits, use Fe(2+) and oxygen to synthesize encapsulated, ferric oxide minerals. Ferritin minerals are iron concentrates stored for cell growth. Ferritins are also antioxidants, scavenging Fenton chemistry reactants. Channels for iron entry and exit consist of helical hairpin segments surrounding the 3-fold symmetry axes of the ferritin nanocages. We now report structural differences caused by amino acid substitutions in the Fe(2+) ion entry and exit channels and at the cytoplasmic pores, from high resolution (1.3-1.8 Å) protein crystal structures of the eukaryotic model ferritin, frog M. Mutations that eliminate conserved ionic or hydrophobic interactions between Arg-72 and Asp-122 and between Leu-110 and Leu-134 increase flexibility in the ion channels, cytoplasmic pores, and/or the N-terminal extensions of the helix bundles. Decreased ion binding in the channels and changes in ordered water are also observed. Protein structural changes coincide with increased Fe(2+) exit from dissolved, ferric minerals inside ferritin protein cages; Fe(2+) exit from ferritin cages depends on a complex, surface-limited process to reduce and dissolve the ferric mineral. High concentrations of bovine serum albumin or lysozyme (protein crowders) to mimic the cytoplasm restored Fe(2+) exit in the variants to wild type. The data suggest that fluctuations in pore structure control gating. The newly identified role of the ferritin subunit N-terminal extensions in gating Fe(2+) exit from the cytoplasmic pores strengthens the structural and functional analogies between ferritin ion channels in the water-soluble protein assembly and membrane protein ion channels gated by cytoplasmic N-terminal peptides.
authors.affiliation: Children's Hospital Oakland Research Institute, Oakland, California 94609, USA.
authors.forename: Takehiko
authors.initials: T
authors.lastname: Tosha
authors.forename: Rabindra K
authors.initials: RK
authors.lastname: Behera
authors.forename: Ho-Leung
authors.initials: HL
authors.lastname: Ng
authors.forename: Onita
authors.initials: O
authors.lastname: Bhattasali
authors.forename: Tom
authors.initials: T
authors.lastname: Alber
authors.forename: Elizabeth C
authors.initials: EC
authors.lastname: Theil
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism; D001001:Anura; D018360:Crystallography, X-Ray; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005290:Ferric Compounds / Q000378:metabolism; D005293:Ferritins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015640:Ion Channel Gating / Q000502:physiology*; D007501:Iron / Q000378:metabolism*; D049329:Nanostructures / Q000737:chemistry*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
chemical_list: D000975:Antioxidants; D005290:Ferric Compounds; C000499:ferric oxide; D005293:Ferritins; D007501:Iron