title: Optimized bacterial production of nonglycosylated human transferrin and its half-molecules.
abstract: 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.
authors.affiliation: Leiden Institute of Chemistry, Department of Biochemistry, Gorlaeus Laboratories, Leiden University, The Netherlands.
authors.forename: M H
authors.initials: MH
authors.lastname: de Smit
authors.forename: P
authors.initials: P
authors.lastname: Hoefkens
authors.forename: G
authors.initials: G
authors.lastname: de Jong
authors.forename: J
authors.initials: J
authors.lastname: van Duin
authors.forename: P H
authors.initials: PH
authors.lastname: van Knippenberg
authors.forename: H G
authors.initials: HG
authors.lastname: van Eijk
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004926:Escherichia coli; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012270:Ribosomes / Q000378:metabolism; D014168:Transferrin / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D010446:Peptide Fragments; D011994:Recombinant Proteins; D014168:Transferrin