Iron binding to human heavy-chain ferritin.
Maxi-ferritins are ubiquitous iron-storage proteins with a common cage architecture made up of 24 identical subunits of five α-helices that drive iron biomineralization through catalytic iron(II) oxidation occurring at oxidoreductase sites (OS). Structures of iron-bound human H ferritin were solved at high resolution by freezing ferritin crystals at different time intervals after exposure to a ferrous salt. Multiple binding sites were identified that define the iron path from the entry ion channels to the oxidoreductase sites. Similar data are available for another vertebrate ferritin: the M protein from Rana catesbeiana. A comparative analysis of the iron sites in the two proteins identifies new reaction intermediates and underlines clear differences in the pattern of ligands that define the additional iron sites that precede the oxidoreductase binding sites along this path. Stopped-flow kinetics assays revealed that human H ferritin has different levels of activity compared with its R. catesbeiana counterpart. The role of the different pattern of transient iron-binding sites in the OS is discussed with respect to the observed differences in activity across the species.
Dipartimento di Biotecnologie, Chimica e Farmacia, Università di Siena, Via Aldo Moro 2, 53100 Siena, Italy.
Cecilia
C
Pozzi
Dipartimento di Biotecnologie, Chimica e Farmacia, Università di Siena, Via Aldo Moro 2, 53100 Siena, Italy.
Flavio
F
Di Pisa
Dipartimento di Chimica, Università di Firenze, Via Della Lastruccia 3, Sesto Fiorentino, 50019 Firenze, Italy.
Caterina
C
Bernacchioni
Dipartimento di Chimica, Università di Firenze, Via Della Lastruccia 3, Sesto Fiorentino, 50019 Firenze, Italy.
Silvia
S
Ciambellotti
Dipartimento di Chimica, Università di Firenze, Via Della Lastruccia 3, Sesto Fiorentino, 50019 Firenze, Italy.
Paola
P
Turano
Dipartimento di Biotecnologie, Chimica e Farmacia, Università di Siena, Via Aldo Moro 2, 53100 Siena, Italy.
Stefano
S
Mangani
D018360:Crystallography, X-Ray; D005293:Ferritins / Q000737:chemistry*; D006801:Humans; D007501:Iron / Q000737:chemistry*; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D011485:Protein Binding
D005293:Ferritins; D007501:Iron
ferritin; human heavy chain; iron; mechanism