Purification and biochemical characterization of recombinant human H-ferritins from Saccharomyces cerevisiae

Abstract

Recombinant human H-ferritins produced from Saccharomyces cerevisiae were purified and their molecular properties were characterized. Electrophoresis of the recombinant H-ferritins showed revealed bands with mobilities similar to those of the H-ferritins produced by Escherichia coli. The pI of H-ferritins from yeast was more basic than that of H-ferritins produced by E. coli. When the cells were treated with tunicamycin, the pI of H-ferritins was driven to a sharp band with the pI range similar to that of those produced by E. coli, implying that the H-ferritins from yeast are glycosylated. The iron uptake of H-ferritins was rapid in the initial stage, with a slight reduction when compared to ferritins from E. coli. Recombinant ferritins are commonly used during synthesis of inorganic nanoparticles in their cores for chemical and industrial purposes. Transmission electron microscopy revealed that the reconstitution yield and size distribution of the core minerals were affected depending on the protein shells. The Hferritins with higher reconstitution yields (64.4%) showed slightly larger sizes (mean 6.52 nm) with narrower size distribution.

Ferritin Nanocages with Biologically Orthogonal Conjugation for Vascular Targeting and Imaging

“…3–16 Human ferritin heavy (hFTH) and light (hFTL) chains are also easily expressed in a variety of microbial expression systems, making their production possible at relatively low cost and large scale. 17–19…”

“…Nature has created a number of molecules with unique architectures and properties suitable for transporting and delivering cargo to specific cells or sites within the body. Among them, the iron-storage protein ferritin is perhaps the strongest candidate for clinical application, given its presence not only within every cell of the human body, but also the extracellular space and circulating plasma. , Ferritin displays remarkable thermal stability (withstanding temperatures up to 80–100 °C), resistance to extreme variation in pH, small size (12 nm in diameter), monodispersity, and a large central cavity for encapsulation of metals, small molecule drugs, contrast agents, and even other nanoparticles. − Human ferritin heavy (hFTH) and light (hFTL) chains are also easily expressed in a variety of microbial expression systems, making their production possible at relatively low cost and large scale. − …”