de Smit M H, Hoefkens P, de Jong G, van Duin J, van Knippenberg P H, van Eijk H G
Leiden Institute of Chemistry, Department of Biochemistry, Gorlaeus Laboratories, Leiden University, The Netherlands.
Int J Biochem Cell Biol. 1995 Aug;27(8):839-50. doi: 10.1016/1357-2725(95)00040-v.
Transferrin is a glycoprotein functioning in iron transport in higher eukaryotes, and consists of two highly homologous domains. To study the function of the glycan residues attached exclusively to the C-terminal domain, we have constructed a plasmid allowing production of nonglycosylated human transferrin in Escherichia coli. By molecular biological and genetic techniques, production was stepped up to 60 mg/l. Similar plasmids were constructed for production of the two half-transferrins. The recombinant proteins accumulate in inclusion-body-like aggregates, where they appear to bind iron without causing bacteriostasis. Proteins active in iron binding have been purified from these inclusion bodies.
