title: Humanized archaeal ferritin as a tool for cell targeted delivery.
abstract: Human ferritins have been extensively studied to be used as nanocarriers for diverse applications and could represent a convenient alternative for targeted delivery of anticancer drugs and imaging agents. However, the most relevant limitation to their applications is the need for highly acidic experimental conditions during the initial steps of particle/cargo assembly, a process that could affect both drug stability and the complete reassembly of the ferritin cage. To overcome this issue the unique assembly of Archaeoglobus fulgidus ferritin was genetically engineered by changing a surface exposed loop of 12 amino acids connecting B and C helices to mimic the sequence of the analogous human H-chain ferritin loop. This new chimeric protein was shown to maintain the unique, cation linked, association-dissociation properties of Archaeoglobus fulgidus ferritin occurring at neutral pH values, while exhibiting the typical human H-homopolymer recognition by the transferrin receptor TfR1. The chimeric protein was confirmed to be actively and specifically internalized by HeLa cells, thus representing a unique nanotechnological tool for cell-targeted delivery of possible payloads for diagnostic or therapeutic purposes. Moreover, it was demonstrated that the 12 amino acids' loop is necessary and sufficient for binding to the transferrin receptor. The three-dimensional structure of the humanized Archaeoglobus ferritin has been obtained both as crystals by X-ray diffraction and in solution by cryo-EM.
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, V.le Regina Elena 291, Rome 00161, Italy. paola.baiocco@iit.it.
authors.forename: Valeria
authors.identifier: http://orcid.org/0000-0003-0872-185X
authors.initials: V
authors.lastname: de Turris
authors.affiliation: Molirom srl, via Ravenna 8, 00161, Rome, Italy.
authors.forename: Matilde
authors.initials: M
authors.lastname: Cardoso Trabuco
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, V.le Regina Elena 291, Rome 00161, Italy. paola.baiocco@iit.it.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Peruzzi
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, V.le Regina Elena 291, Rome 00161, Italy. paola.baiocco@iit.it and Institute of Molecular Biology and Pathology, National Research Council, P.le A. Moro, 7, 00185, Rome, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Boffi
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, V.le Regina Elena 291, Rome 00161, Italy. paola.baiocco@iit.it.
authors.forename: Claudia
authors.initials: C
authors.lastname: Testi
authors.affiliation: Institute of Molecular Biology and Pathology, National Research Council, P.le A. Moro, 7, 00185, Rome, Italy.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Vallone
authors.affiliation: Department of Biochemical Sciences "Alessandro Rossi Fanelli", Sapienza University of Rome, P.le A. Moro, 5, 00185, Rome, Italy.
authors.forename: Linda
authors.initials: L
authors.lastname: Celeste Montemiglio
authors.affiliation: The City University of New York Advanced Science Research Center 85 St. Nicholas Terrace, New York, NY 10031, USA.
authors.forename: Amédée Des
authors.identifier: http://orcid.org/0000-0002-9704-3781
authors.initials: AD
authors.lastname: Georges
authors.affiliation: Department of Biochemical Sciences "Alessandro Rossi Fanelli", Sapienza University of Rome, P.le A. Moro, 5, 00185, Rome, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Calisti
authors.affiliation: Department of Biochemical Sciences "Alessandro Rossi Fanelli", Sapienza University of Rome, P.le A. Moro, 5, 00185, Rome, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Benni
authors.affiliation: Department of Biochemical Sciences "Alessandro Rossi Fanelli", Sapienza University of Rome, P.le A. Moro, 5, 00185, Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Bonamore
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, V.le Regina Elena 291, Rome 00161, Italy. paola.baiocco@iit.it.
authors.forename: Paola
authors.identifier: http://orcid.org/0000-0002-1084-5694
authors.initials: P
authors.lastname: Baiocco
mesh_terms: D015703:Antigens, CD / Q000737:chemistry; D001052:Apoferritins / Q000737:chemistry; D019611:Archaeoglobus fulgidus / Q000737:chemistry*; D016503:Drug Delivery Systems; D005293:Ferritins / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D015202:Protein Engineering; D011990:Receptors, Transferrin / Q000737:chemistry
chemical_list: D015703:Antigens, CD; C068126:CD71 antigen; D011990:Receptors, Transferrin; D005293:Ferritins; D001052:Apoferritins